Aminoquinoline derivatives and their use as adenosine A3 ligands

ABSTRACT

The present invention relates to an adenosine A 3  receptor ligand of the general formula (I), within those preferably to the antagonists, including a salt, solvate or isomer (tautomer, desmotrop, and optically active isomer) thereof, to a pharmaceutical composition containing the ligand, to the use of the ligand, to its preparation, and intermediates of the ligand of the general formula (II″), (III″), (IV″), (V″), (VI″), (VII″), (VIII″) and (XIII″) and their preparation.

CROSS REFERENCE

This application is a continuation of PCT Application HU2004/000080,filed 23 Jul. 2004, which designated the United States.

FIELD OF THE INVENTION

The present invention relates to an adenosine A₃ receptor ligand of thegeneral formula (I), within those preferably to the antagonists,including a salt, solvate or isomer (tautomer, desmotrop, and opticallyactive isomer) thereof, to a pharmaceutical composition containing theligand, to the use of the ligand, to its preparation, and intermediatesof the ligand of the general formula (II″), (III″), (IV″), (V″), (VI″),(VII″), (VIII″) and (XIII″) and their preparation.

BACKGROUND OF THE INVENTION

Adenosine is a well-known component of several endogenous molecules(ATP, NAD⁺, nucleic acids). It plays an important regulatory role inmany physiological processes. The effect of adenosine on the heartfunction was described already in 1929 (Drury and Szentgyörgyi, Physiol.68:213, 1929). The identification of an increasing number ofphysiological functions mediated by adenosine and the discovery of newadenosine receptor subtypes are offering possibilities for thetherapeutic application of specific ligands (Poulse, S. A. and Quinn, R.J. Bioorganic and Medicinal Chemistry 6:619, 1998).

To date, the receptors for adenosine have been classified into threemain classes: A₁, A₂ and A₃. The A₁ subtype is partly responsible forthe inhibition of the adenylate cyclase by coupling to G_(i) membraneprotein, and partly influences other second messenger systems. The A₂receptor subtype can be subdivided into two further subtypes—A_(2a) andA_(2b)—which stimulate the adenylate cyclase activity. The sequence ofthe adenosine A₃ receptors have been first identified from rat testiscDNA library. Later it was proved that it corresponds to a novel,functional adenosine receptor. The activation of the A₃ receptors isalso connected with several second-messenger systems: inhibiting ofadenylate cyclase, stimulating phospholipase C and D.

The adenosine receptors are found in several organs and regulate theirfunctions. Both A₁ and A_(2a) receptors play important role in thecentral nervous system and cardiovascular system. In the CNS, theadenosine inhibits the release of synaptic transmitters which effect ismediated by A₁ receptors. In the heart, the A₁ receptors also mediatethe negative inotropic, chronotropic and dromotropic effects ofadenosine. The adenosine A_(2a) receptors, which are located in arelatively high amount in the striatum, display functional interactionwith the dopamine receptors in regulating the synaptic transmission. TheA_(2a) adenosine receptors on endothelial and smooth muscle cells areresponsible for adenosine-induced vasodilation.

On the basis of RNA identification, the A_(2b) adenosine receptors arewidely distributed in different tissues. They have been identified inalmost every cell type, but its expression is the highest in theintestine and the bladder. This subtype probably also has importantregulatory function in the regulation of the vascular tone and plays arole in the function of mast cells.

Contrary to A₁ and A_(2a) receptors, where the tissue distribution wasdetected on the protein level, the presence of A_(2b) and A₃ receptorswas detected on the basis of their mRNA level. Expression levels for A₃adenosine receptors are rather low compared to other subtypes and theyare highly species dependent. A₃ adenosine receptors are expressedprimarily in the central nervous system, in the testis and in the immunesystem, and appear to be involved in the modulation of the mediatorrelease from the mast cells in immediate hypersensitivity reaction.

For therapeutic use, it is essential to ensure that the molecule doesnot bind, or binds only in the case of very high concentration to theA₁, A_(2a) and A_(2b) sub-types of the adenosine receptor.

A₃ antagonists published so far in the literature, belong to the groupsof flavonoides, 1,4-dihydropyridine derivatives, triazoloquinazolines,thiazolonaphthyridines and thiazolopyrimidines. Most of the effectiveand for the adenosine subtypes selective antagonists, however possesstrong lipophilic character, and they are therefore sparingly soluble inwater. This feature hinders the in vivo applicability of the compounds.In the literature more and more studies are to find aiming thepreparation of water-soluble adenosine A3 receptor antagonists (Ch. E.Müller et al., J. Med. Chem. 45:3440, 2002; A. Maconi et al., J. Med.Chem. 45:3579, 2002).

Patent application WO 02/096879 discloses 2-amino-3-cyanoquinolinederivatives as structurally novel type, effective A₃ antagonists. Thecompounds described in patent application WO 02/096879 are A₃antagonists with high selectivity of the following general formula:

-   R^(1′) stands for hydrogen atom or straight or branched C₁₋₄ alkyl    group;-   R^(2′) stands for hydrogen atom or straight or branched C₁₋₄ alkyl    group;-   R^(3′) stands for hydrogen atom or straight or branched C₁₋₄ alkyl    group, phenyl, thienyl, or furyl group, optionally substituted with    one or more straight or branched C₁₋₄ alkyl group, straight or    branched C₁₋₄ alkoxy group or halogen atom; a six- or five-membered    heteroaromatic ring containing one, two or three nitrogen atoms, or    one nitrogen atom and one oxygen atom, or one nitrogen atom and one    sulphur atom, optionally substituted with one or more straight or    branched C₁₋₄ alkyl group, straight or branched C₁₋₄ alkoxy group or    halogen atom;-   R^(9′), R^(10′), R^(11′), and R^(12′) independently stand for    hydrogen atom, straight or branched C₁₋₄ alkyl group, straight or    branched C₁₋₄ alkoxy group, hydroxy group or halogen atom, or R^(9′)    and R^(12′) stand for hydrogen atom and R^(10′) and R^(11′) form    together a methylenedioxy group;-   R^(6′) stands for hydrogen atom or a cyano group, aminocarbonyl    group, C₁₋₄ alkoxycarbonyl group, or carboxy group;-   R^(7′) stands for hydrogen atom or straight or branched C₁₋₄ alkyl    group, phenyl, benzyl, thienyl, or furyl group, optionally    substituted with methylenedioxy-group or with one or more straight    or branched C₁₋₄ alkyl group, straight or branched C₁₋₄ alkoxy    group, hydroxy group, trifluoromethyl group, cyano group or halogen    atom; or a six- or five-membered heteroaromatic ring containing one,    two or three nitrogen atoms, or one nitrogen atom and one oxygen    atom, or one nitrogen atom and one sulphur atom, optionally    substituted with one or more straight or branched C₁₋₄ alkyl group,    straight or branched C₁₋₄ alkoxy group or halogen atom;-   X′ stands for —CH₂— group, —NH— group, —NR^(8′)— group, or sulphur    atom, oxygen atom, sulpho group or sulphoxy group, wherein R^(8′)    stands for straight or branched C₁₋₄ alkyl group or C₃₋₆ cycloalkyl    group;-   n′ represents zero, 1 or 2;

These compounds too, have the characteristic disadvantage that they areonly sparingly soluble, which hampers their development into a drug.What would be useful would be compounds having solubility profiles thatare far better than those of the known 2-amino-3-cyanoquinolines,besides, and which would also be highly active.

SUMMARY OF THE INVENTION

The instant invention is directed to a compound of the general formula(I),

wherein

-   R¹ stands for hydrogen atom or a straight or branched C₁₋₄ alkyl;-   R² stands for hydrogen atom or a straight or branched C₁₋₄ alkyl;-   R³ stands for hydrogen atom, straight or branched C₁₋₄ alkyl, or    C₃₋₆ cycloalkyl, phenyl, thienyl, or furyl which is optionally    substituted with one or more straight or branched C₁₋₄ alkyl,    straight or branched C₁₋₄ alkoxy, or halogen atom, or six- or    five-membered heteroaromatic ring containing one, two or three    nitrogen atoms, or five-membered heteroaromatic ring containing one    nitrogen atom and one oxygen atom, or one nitrogen atom and one    sulphur atom, which heteroaromatic ring is optionally substituted    with one or more straight or branched C₁₋₄ alkyl, straight or    branched C₁₋₄ alkoxy, or halogen atom;-   R⁴ and R⁵ independently stand for hydrogen atom, C₃₋₆ cycloalkyl,    straight or branched C₁₋₄ alkyl which is optionally substituted by a    hydroxy, carboxy, or straight or branched C₁₋₄ alkoxy, amino or    amino substituted with one or two straight or branched C₁₋₄ alkyl;    or-   R⁴ stands for hydrogen atom, straight or branched C₁₋₄ alkyl or    benzyl, and-   R⁵ stands for hydrogen atom, —SO₂OH or straight or branched C₁₋₄    acyl, or-   R⁴ and R⁵ taken together with the nitrogen atom to which they are    attached form a group of the general formula a.)

-   R⁶ stands for hydrogen atom or straight or branched C₁₋₄ alkyl,    phenyl, benzyl, thienyl, or furyl, each of which is optionally    substituted with methylenedioxy, or one or more straight or branched    C₁₋₄ alkyl, straight or branched C₁₋₄ alkoxy, hydroxy,    trifluoromethyl, cyano or halogen atom, or six- or five-membered    heteroaromatic ring containing one, two or three nitrogen atoms, or    one nitrogen atom and one oxygen atom, or one nitrogen atom and one    sulphur atom, which heteroaromatic ring is optionally substituted    with one or more straight or branched C₁₋₄ alkyl, straight or    branched C₁₋₄ alkoxy or halogen atom;-   R⁷ and R⁸ independently stand for hydrogen atom, straight or    branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl;-   X stands for —CH₂—, —NH—, —NR⁹—, or sulphur atom, oxygen atom,    sulpho or sulphoxy;-   R⁹ stands for straight or branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl;-   Z stands for oxygen atom, sulphur atom, —CHR¹⁰— or —NR¹¹—;-   R¹⁰ stands for hydrogen atom, straight or branched C₁₋₄ alkyl, or    C₃₋₆ cycloalkyl;-   R¹¹ stands for hydrogen atom, straight or branched C₁₋₄ alkyl, C₃₋₆    cycloalkyl, —SO₂OH or straight or branched C₁₋₄ acyl;-   n represents zero, 1 or 2;-   m represents 1, 2, or 3;-   o represents 1, 2, or 3;-   p represents zero or 1; and-   r represents zero or 1, or    a salt, solvate, or isomer (tautomer, desmotrop, optically active    isomer) thereof, or a salt or solvate of the isomer. The compound of    the general formula (I) have a high selectivity for the A₃ sub-type    of the adenosine receptor.

DETAILED DESCRIPTION OF THE INVENTION

Our aim was to prepare A₃ ligands, within them preferably antagonists,with quinoline structure, which exert strong antagonistic effect andhigh selectivity for the A₃ receptor, i.e. they inhibit the A₃ receptorin much lower concentration than they inhibit the A₁, A_(2a) and A_(2b)receptors. Further aims were to have stability, bioavailability,therapeutic index, toxicity and solubility data, which enable these newcompounds to develop into drug substances, and that the new compoundspossess favourable enteric absorption to be applied orally.

As shown by liquid phase NMR studies, in solution the compound of thegeneral formula (I) is in equilibrium mixtures of the tautomeric forms(IA) and (IB):

In solid phase the tautomers (IA) and (IB) can be isolated separately,as the desmotrop (IA) or (IB).

According to the above findings, the present invention also relates tothe desmotropic isomers (IA) and (IB). The meanings of the substituentsin the general formulae (IA) and (IB) are as defined for the generalformula (I).

The compound of the general formula (I) may have a chiral centredepending for instance on the meaning of the set of substituents R¹, R²,and R³. Thus, the invention also relates to the racemic form or anoptically active form of the compound of the general formula (I), or asalt, tautomer or desmotrop thereof.

Definition of Terms

Detailed meanings of the above substituents are as follows:

By a straight or branched C₁₋₄ alkyl we mean a methyl, ethyl, propyl,isopropyl, butyl, isobutyl, secondary-butyl or tertiary-butyl,preferably ethyl or methyl.

By a straight or branched C₁₋₄ alkoxy we mean a methoxy, ethoxy,propoxy, isopropoxy, butoxy, isobutoxy, secondary-butoxy,tertiary-butoxy, preferably ethoxy or methoxy.

By a C₃₋₆ cycloalkyl we mean a cyclopropyl, cyclobutyl, cyclopentyl orcyclohexyl.

By a straight or branched C₁₋₄ acyl we mean a formyl, acetyl, propionyl,2-methyl-propionyl, or butyryl.

By the heteroaromatic ring containing one, two or three nitrogen atomswe mean a pyrrole, imidazole, pyrazole, 1,2,3-triazole, 1,2,4-triazole,pyridine, pyrimidine, pyridazine, pyrazine or 1,2,4-triazine ring. Thering may optionally be substituted with a C₁₋₄ alkyl, C₁₋₄ alkoxy, orhalogen atom.

By the heteroaromatic ring containing one nitrogen atom and one oxygen-or one sulphur atom we mean an oxazole, isoxazole, thiazole orisothiazole ring. The ring may optionally be substituted with a C₁₋₄alkyl, C₁₋₄ alkoxy, or halogen atom.

The a.) group

preferably represents a pyrrolidino, piperidino, piperazino,4-methylpiperazino, 4-formylpiperazino, 4-sulfonylpiperazino ormorpholino.

By a salt of the compound of the general formula (I) we mean a saltformed with inorganic or organic acids and bases. Preferred salts aregiven with pharmaceutically accepted acids, as for instance hydrochloricacid, sulphuric acid, ethanesulphonic acid, tartaric acid, malic acid,citric acid, fumaric acid, and with pharmaceutically accepted bases, asfor instance NaOH, potassium hydroxide, and ethanolamine. Salts used forpurification or isolation, as for instance the methanesulphonate ortetrafluoroborate salts are also subjects of the invention.

By a solvate we mean a solvate of various solvents, as for instance ofwater or ethanol.

By an isomer we mean a tautomer, desmotrop, or optically active isomer.

Particular or Preferred Embodiments of the Invention

A favoured embodiment of the compound of the general formula (I) iswherein

-   R¹ stands for hydrogen atom or a straight or branched C₁₋₄ alkyl;-   R² stands for hydrogen atom or a straight or branched C₁₋₄ alkyl;-   R³ stands for hydrogen atom, a straight or branched C₁₋₄ alkyl or    C₃₋₆ cycloalkyl, a phenyl, thienyl, or furyl, each of which is    optionally substituted with one or more straight or branched C₁₋₄    alkyl, straight or branched C₁₋₄ alkoxy, or halogen atom;-   R⁴ and R⁵ independently stand for hydrogen atom, C₃₋₆ cycloalkyl,    straight or branched C₁₋₄ alkyl which is optionally substituted by a    hydroxy, carboxy, straight or branched C₁₋₄ alkoxy, amino, or amino    substituted with one or two straight or branched C₁₋₄ alkyl; or-   R⁴ stands for hydrogen atom or a straight or branched C₁₋₄ alkyl or    a benzyl, and-   R⁵ stands for hydrogen atom, —SO₂OH or a straight or branched C₁₋₄    acyl, or-   R⁴ and R⁵ taken together with the nitrogen atom to which they are    attached form a group of the general formula a.)

-   R⁶ stands for hydrogen atom or straight or branched C₁₋₄ alkyl,    phenyl, benzyl, thienyl, or furyl, each of which is optionally    substituted with methylenedioxy, or with one or more straight or    branched C₁₋₄ alkyl, straight or branched C₁₋₄ alkoxy, hydroxy,    trifluoromethyl, cyano, or halogen atom; or six- or five-membered    heteroaromatic ring containing one, two or three nitrogen atoms, or    one nitrogen atom and one oxygen atom, or one nitrogen atom and one    sulphur atom, which heteroaromatic ring is optionally substituted    with one or more straight or branched C₁₋₄ alkyl, straight or    branched C₁₋₄ alkoxy or halogen atom;-   R⁷ and R⁸ independently stand for hydrogen atom, straight or    branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl;-   X stands for —CH₂—, —NH—, —NR⁹—, or sulphur atom, oxygen atom,    sulpho or sulphoxy;-   R⁹ represents a straight or branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl;-   Z stands for oxygen atom, sulphur atom, —CHR¹⁰— or —NR¹¹—;-   R¹⁰ represents hydrogen atom, straight or branched C₁₋₄ alkyl, or    C₃₋₆ cycloalkyl;-   R¹¹ stands for hydrogen atom, straight or branched C₁₋₄ alkyl, C₃₋₆    cycloalkyl, —SO₂OH or formyl;-   n represents zero, 1 or 2;-   m represents 1, 2, or 3;-   o represents 1, 2, or 3;-   p represents zero or 1; and-   r represents zero or 1, or    a salt, solvate, or isomer (tautomer, desmotrop, optically active    isomer) thereof, or a salt or solvate of the isomer.

A special embodiment of the compound of the general formula (I) iswherein

-   R¹ stands for hydrogen atom or methyl;-   R² stands for hydrogen atom or methyl;-   R³ stands for phenyl, thienyl, or furyl;-   R⁴ and R⁵ independently stand for hydrogen atom, C₃₋₆ cycloalkyl,    straight or branched C₁₋₄ alkyl which is optionally substituted by a    hydroxy, carboxy, or straight or branched C₁₋₄ alkoxy, amino, or    amino substituted with one or two straight or branched C₁₋₄ alkyl,    or-   R⁴ stands for hydrogen atom or a straight or branched C₁₋₄ alkyl or    a benzyl, and-   R⁵ stands for hydrogen atom, —SO₂OH or a straight or branched C₁₋₄    acyl, or-   R⁴ and R⁵ taken together with the nitrogen atom to which they are    attached form a group of the general formula a.),

-   R⁶ stands for 4-methoxyphenyl, 3-methylphenyl, 3-methoxyphenyl,    3,4-methylene-dioxyphenyl, 4-fluorophenyl, 2-thienyl or 2-furyl;-   R⁷ and R⁸ independently stand for hydrogen atom, straight or    branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl;-   X stands for —NH—, or oxygen atom;-   Z stands for oxygen atom, sulphur atom, —CH₂—, —NH— or —NR¹¹—-   R¹¹ represents a straight or branched C₁₋₄ alkyl, C₃₋₆ cycloalkyl,    —SO₂OH or formyl; and-   n represents 1;-   m represents 2;-   o represents 2;-   p represents zero; and-   r represents zero, or    a salt, solvate, or isomer (tautomer, desmotrop, optically active    isomer) thereof, or a salt or solvate of the isomer.

Especially favoured is a compound selected from the following:

-   4-Methoxy-N-[6-(morpholin-4-yl)-4-benzylamino-3-cyanoquinolin-2-yl]-benzamide,-   4-Methoxy-N-[6-(4-methylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide,-   4-Methoxy-N-(6-dimethylamino-4-benzylamino-3-cyanoquinolin-2-yl)benzamide,-   3,4-Methylenedioxy-N-(6-dimethylamino-4-benzylamino-3-cyanoquinolin-2-yl)benzamide,-   4-Fluoro-N-(6-dimethylamino-4-benzylamino-3-cyanoquinolin-2-yl)benzamide,-   4-Methoxy-N-(6-(piperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl)benzamide,-   4-Methoxy-N-(6-amino-4-benzylamino-3-cyanoquinolin-2-yl)benzamide,-   N-[4-(Benzylamino)-3-cyano-6-(4-formylpiperazin-1-yl)quinolin-2-yl]-4-methoxybenzamide,-   4-{4-(Benzylamino)-3-cyano-2-[(4-methoxybenzoyl)amino]quinolin-6-yl}piperazine-1-sulfonic    acid,-   N-{3-Cyano-6-(formylamino)-4-[(2-thienylmethyl)amino]quinolin-2-yl}-4-methoxybenzamide,-   N-{3-Cyano-6-(formylamino)-4-[(2-thienylmethyl)amino]quinolin-2-yl}-1,3-benzodioxole-5-carboxamide,-   N-[4-(Benzylamino)-3-cyano-6-(formylamino)quinolin-2-yl]-1,3-benzodioxole-5-carboxamide,-   N-[4-(Benzylamino)-3-cyano-6-(formylamino)quinolin-2-yl]-4-methoxybenzamide,-   N-{4-(Benzylamino)-3-cyano-6-[formyl(methyl)amino]quinolin-2-yl}-4-methoxybenzamide,-   N-{3-Cyano-6-[formyl(methyl)amino]-4-[(2-thienylmethyl)amino]quinolin-2-yl}-4-methoxybenzamide,-   {3-Cyano-2-[(4-methoxybenzoyl)amino]-4-[(2-thienylmethyl)amino]quinolin-6-yl}methylsulfamic    acid,-   {4-(Benzylamino)-3-cyano-2-[(4-methoxybenzoyl)amino]quinolin-6-yl}methylsulfamic    acid,-   {4-(Benzylamino)-3-cyano-2-[(4-methoxybenzoyl)amino]quinolin-6-yl}sulfamic    acid,-   [2-[(1,3-Benzodioxol-5-ylcarbonyl)amino]-4-(benzylamino)-3-cyanoquinolin-6-yl]sulfamic    acid,-   {2-[(1,3-Benzodioxol-5-ylcarbonyl)amino]-3-cyano-4-[(2-thienylmethyl)amino]quinolin-6-yl}sulfamic    acid, and-   {3-Cyano-2-[(4-methoxybenzoyl)amino]-4-[(2-thienylmethyl)amino]quinolin-6-yl}sulfamic    acid, or    a salt, solvate, or isomer (tautomer, desmotrop, optically active    isomer) thereof, or a salt or solvate of the isomer.

The compound of the general formula (I) according to the presentinvention as well as a salt, solvate, or isomer (tautomer, desmotrop,optically active isomer) thereof, or a salt or solvate of the isomer,displays suitable solubility, therefore can favourably be used as anactive substance in a pharmaceutical composition.

Solubility values in water and in aqueous buffer solutions (pH=6.5 and7.5) for the compound of the formula (1) of patent application WO02/096879 are less than 1 mg/L, whereas those for the compound of thegeneral formula (I) of the present invention are between 1-200 mg/L. Thepresent invention also relates to a pharmaceutical compositioncontaining as an active principle the compound of the general formula(I) or a salt, solvate, or isomer (tautomer, desmotrop, optically activeisomer) thereof, or a salt or solvate of the isomer, which is preferablyan oral composition, but an inhalable, parenteral and transdermalformulation is also an aspect of the invention. The above pharmaceuticalcompositions may be solids or liquids, such as tablets, pellets,capsules, patches, solutions, suspensions or emulsions. Solidcompositions, first of all tablets and capsules are preferred.

The above pharmaceutical compositions are prepared by applying usualpharmaceutical auxiliary materials and by using standard methods.

The compound of the general formula (I) can be used for the treatment ofpathologies, where A₃ receptor plays a role in the development of thedisease.

The compound of the present invention having selective activity on theA₃ receptor can be used in the therapeutic and/or preventive treatmentof disfunctions of the heart, kidney, respiratory system, centralnervous system. It inhibits the protective effect of adenosine ingrowing tumor cells, prevent mast cell degranulation, inhibits thecytokine production, reduce the inraocular pressure, inhibits the TNFαrelease, inhibits the migration of eosinophils, neutrophils and otherimmune cells, and inhibits the bronchoconstriction and plasmaextravasation.

Based on these effects, adenosine A₃ receptor antagonists of the presentinvention may be therapeutically useful as antiinflammatory,antiasthmatic, antiischemic, antidepressant, antiarrhytmic, renalprotective, antitumor, antiparkinson and cognitive enhancing drugs. Theyalso may be useful in the treatment or prevention of miocardialreperfusion injury, chronic obstructive pulmonary disease (COPD) andadult respiratory distress syndrome (ARDS) including chronic bronchitis,pulmonary emphysema or dyspnea, allergic reactions (e.g. rhinitis,poison ivy induced responses, urticaria, scleroderma, arthritis) otherautoimmune diseases, inflammatory bowel disease, Addison's disease,Crohn's disease, psoriasis, rheumatism, hypertension, neurogicalfunction disorders, glaucoma and diabetes (K. N. Klotz,Naunyn-Schmiedberg's Arch. Pharmacol. 362:382, 2000; P. G. Baraldi és P.A. Borea, TiPS 21:456, 2000).

The compound of the present invention may be preferably used for themanufacture of a pharmaceutical composition for the treatment ofdiseases such as asthma, COPD and ARDS, glaucoma, tumor, allergic andinflammatory diseases, ischemia, hypoxia, arrythmia and renal diseases.

The present invention relates furthermore to the use of the compound ofthe general formula (I) in the treatment of the above pathologies. Thesuggested daily dose is 0.1-1000 mg active ingredient, depending on thenature and severeness of the disease and on the sex, weight etc. of thepatient.

A further aspect of the invention is the preparation of the compound ofthe general formula (I).

The substituents in the formulae of the intermediates and reagents ofthe general formulae (I″), (II″), (III″), (IV″), (V″), (VI″), (VII″),(VIII″), (IX″), (X″), (XI″), (XII″), (XIII″) and (XIV″):

have the meanings as follows.

-   R^(1″) stands for hydrogen atom or a straight or branched C₁₋₄    alkyl;-   R^(2″) stands for hydrogen atom or a straight or branched C₁₋₄    alkyl;-   R^(3″) stands for hydrogen atom, straight or branched C₁₋₄ alkyl, or    C₃₋₆ cycloalkyl, phenyl, thienyl, or furyl, each of which is    optionally substituted with one or more straight or branched C₁₋₄    alkyl, straight or branched C₁₋₄ alkoxy, or halogen atom; six- or    five-membered heteroaromatic ring containing one, two or three    nitrogen atoms, or five-membered heteroaromatic ring containing one    nitrogen atom and one oxygen atom, or one nitrogen atom and one    sulphur atom, which heteroaromatic ring is optionally substituted    with one or more straight or branched C₁₋₄ alkyl, straight or    branched C₁₋₄ alkoxy, or halogen atom;-   R^(4″) and R^(5″) independently stand for hydrogen atom, C₃₋₆    cycloalkyl, straight or branched C₁₋₄ alkyl which is optionally    substituted by a hydroxy, a carboxy, or straight or branched C₁₋₄    alkoxy, amino, amino substituted with one or two straight or    branched C₁₋₄ alkyl, or protective group;-   R^(4″) stands for hydrogen atom or a straight or branched C₁₋₄ alkyl    or a benzyl, and-   R^(5″) stands for hydrogen atom, —SO₂OH, straight or branched C₁₋₄    acyl or protective group, or-   R^(4″) and R^(5′) taken together with the nitrogen atom to which    they are attached form a group of the general formula a″.)

-   R^(6″) stands for hydrogen atom or straight or branched C₁₋₄ alkyl,    phenyl, benzyl, thienyl, or furyl, each of which is optionally    substituted with methylenedioxy, or one or more straight or branched    C₁₋₄ alkyl, straight or branched C₁₋₄ alkoxy, hydroxy,    trifluoromethyl, cyano or halogen atom; or six- or five-membered    heteroaromatic ring containing one, two or three nitrogen atoms, or    one nitrogen atom and one oxygen atom, or one nitrogen atom and one    sulphur atom, which heteroaromatic ring is optionally substituted    with one or more straight or branched C₁₋₄ alkyl, straight or    branched C₁₋₄ alkoxy or halogen atom;-   R^(7″) and R^(8″) independently stand for hydrogen atom, straight or    branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl;-   X″ stands for —CH₂—, —NH—, —NR^(9″)—, or sulphur atom, oxygen atom,    sulpho or sulphoxy;-   R^(9″) stands for straight or branched C₁₋₄ alkyl or C₃₋₆    cycloalkyl;-   Z″ stands for oxygen atom sulphur atom, —CHR^(10 ″)— or —NR^(11 ″)—;-   R^(10″) stands for hydrogen atom, straight or branched C₁₋₄ alkyl,    or C₃₋₆ cycloalkyl;-   R^(11″) stands for hydrogen atom, straight or branched C₁₋₄ alkyl,    C₃₋₆ cycloalkyl, —SO₂OH, straight or branched C₁₋₄ acyl or    protective group;-   R^(12″) stands for straight or branched C₁₋₃ alkyl;-   Y″ represents leaving group;-   n″ represents zero, 1 or 2;-   m″ represents 1, 2, or 3;-   o″ represents 1, 2, or 3;-   p″ represents zero or 1;-   r″ represents zero or 1.

A further aspect of the invention is the preparation of the compound ofthe general formula (I) and of the partly novel intermediates of thegeneral formulae (I″), (II″), (III″), (IV″), (V″), (VI″), (VII″),(VIII″), and (XIII″).

In the process aspects according to the invention

v.) the bis-carboxamide of the general formula (II″) is selectivelyhydrolysed and if desired the protective group removed, or

v/i.) for the preparation of a compound of the general formula (I),wherein R⁴ stands for hydrogen atom, straight or branched C₁₋₄ alkyl orbenzyl and R⁵ stands for straight or branched C₁₋₄ acyl, or R⁴ and R⁵taken together with the nitrogen atom to which they are attached form agroup of the general formula a.), wherein Z stands for a —NR¹¹, whereR¹¹ stands for a straight or branched C₁₋₄ acyl and R¹, R², R³, R⁶, R⁷,R⁸, R⁹, X, n, m, o, p and r are as defined above,

a compound of the general formula (I), wherein R⁴ stands for hydrogenatom, a straight or branched C₁₋₄ alkyl or benzyl and R⁵ stands forhydrogen atom, or R⁴ and R⁵ together with the nitrogen atom form a groupof the general formula a.), wherein Z stands for —NH and R¹, R², R³, R⁶,R⁷, R⁸, R⁹, X, n, m, o, p and r are as defined above,

is acylated with a compound of the general formula R^(12″)COY″, whereinR^(12″) and Y″ are as defined above, or

v/ii.) for the preparation of a compound of the general formula (I),wherein R⁴ stands for hydrogen atom, a straight or branched C₁₋₄ alkylor benzyl and R⁵ stands for —SO₂OH, or R⁴ and R⁵ taken together with thenitrogen atom to which they are attached form a group of the generalformula a.), wherein Z stands for —NR¹¹—, where R¹¹ stands for —SO₂OHand R¹, R², R³, R⁶, R⁷, R⁸, R⁹, X, n, m, o, p and r are as definedabove,

a compound of the general formula (I), wherein R⁴ stands for hydrogenatom, a straight or branched C₁₋₄ alkyl or benzyl and R⁵ stands forhydrogen atom, or R⁴ and R⁵ together with the nitrogen atom form a groupof the general formula a.), wherein Z stands for —NH— and R¹, R², R³,R⁶, R⁷, R⁸, R⁹, X, n, m, o, p and r are as defined above,

is reacted with a pyridine-SO₃ complex, or with ClSO₃H, or

v/iii.) for the preparation of a compound of the general formula (I),wherein R⁴ stands for hydrogen atom, a straight or branched C₁₋₄ alkylor benzyl and R⁵ stands for —SO₂OH and R¹, R², R³, R⁶, R⁷, R⁸, R⁹, X, Z,n, m, o, p and r are as defined above,

a compound of the general formula (XIII″), wherein R¹, R², R³, R⁶, R⁹,X, and n are as defined above

is reacted with Na₂S₂O₄ or NaHSO₃, or

is reduced and the compound of the general formula (XIV″), thusobtained, wherein R^(4″) stands for hydrogen atom and R^(1″), R^(2″),R^(3″), R^(6″), R^(9″), X″, and n″, are as defined above, is sulphatedwith a pyridine-SO₃ complex or with ClSO₃H,

if desired after transforming the R^(4″) hydrogen atom into straight orbranched C₁₋₄ alkyl or benzyl, or

v/iv.) for the preparation of a compound of the general formula (I),wherein R⁴ stands for hydrogen atom, C₃₋₆ cycloalkyl, benzyl, straightor branched C₁₋₄ alkyl which is optionally substituted by hydroxy,carboxy, or straight or branched C₁₋₄ alkoxy, amino, or aminosubstituted with one or two straight or branched C₁₋₄ alkyl, and R⁵stands for hydrogen atom, or

R⁴ and R⁵, taken together with the nitrogen atom to which they areattached form a group of the general formula a.), wherein R⁷ and R⁸independently stand for hydrogen atom, straight or branched C₁₋₄ alkylor C₃₋₆ cycloalkyl and Z stands for —NR¹¹, where R¹¹ stands for hydrogenatom,

removing the protective group of a compound of the general formula (I″),wherein R^(4″) stands for hydrogen atom, C₃₋₆ cycloalkyl, benzyl,straight or branched C₁₋₄ alkyl which is optionally substituted byhydroxy, a carboxy, or straight or branched C₁₋₄ alkoxy, amino, or aminosubstituted with one or two straight or branched C₁₋₄ alkyl, and R^(5″)stands for a protective group, or R^(4″) and R^(5″), taken together withthe nitrogen atom to which they are attached form a group of the generalformula a″.), wherein R^(7″) and R^(8″) independently stand for hydrogenatom, straight or branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl and Z standsfor —NR^(11″), where R^(11″) stands for a protecting group,

and—if desired—the substituents of the resulting compound of the generalformula (I) are transformed into each other by known methods, and/or thecompound of the general formula (I) thus obtained is transformed intoits salt or solvate, or liberated from its salt or solvate and—ifdesired—resolved into its optically active isomers and—if desired—agiven desmotrop is transformed into an other desmotropic form.

As for agents for the selective hydrolysis, alkali hydroxides,preferably potassium hydroxide and/or NaOH dissolved in an alcohol,preferably in methanol can be used, but other agents known in theorganic chemistry helping the amide hydrolysis, can also be applied.

The selective hydrolysis can be carried out in a broad temperaturerange, preferable is the range between 20° C. and 100° C.

The reaction v/i.) can be performed using an ester derivative of theappropriate acid at elevated temperature or using an activated acidderivative (e.g. acid-halogenid) at room temperature or elevatedtemperature applying a suitable base (e.g. triethylamine).

The reaction v/ii.) can be performed using pyridine-SO₃ complex in asuitable solvent (preferably in pyridine) or using ClSO₃H in a suitablesolvent (preferably in pyridine or chloroform) applying a strong organicbase (like DBU) or an inorganic base (like K₂CO₃) at room temperature orelevated temperature.

The reaction v/iii.) can be performed in an appropriate aqueous alkalinesolution of the nitro compound using an excess of Na₂S₂O₄ or NaHSO₃ asreagent at room temperature or at elevated temperature (100°).

According to a variation of the reaction v/iii.) the nitro derivative isreduced by any of the known methods and the resulting amino derivativeis alkylated or benzylated first under known reductive alkylation orbenzylation conditions and the alkylamino or benzylamino derivatives,thus obtained will be sulphated as described under ii.).

The protective group may be any known protective group. Examples ofprotecting groups, as well as methods for protecting and deprotectingvarious functional groups are given in “Protective Groups in OrganicSynthesis”, Green et al., 2^(nd) Edition *John Wiley & Sons, Inc, NewYork., 1991. Preferred protective groups are Boc, ethoxycarbonyl,benzyloxycarbonyl, etc, most preferably benzyl group.

The substituents of the compound of the general formula (I) can betransformed into each other by known methods (Comprehensive OrganicTransformation, R. C. Larock, VCH Publisher, New York, 1989).

The salts can be prepared by adding one equivalent of the acid to thealcoholic, preferably ethanol solution of the base and diluting thesolution with diethyl ether and finally filtering off and drying theresulting crystals.

The compound of the general formula (II″)—wherein the meanings ofR^(1″), R^(2″), R^(3″), R^(4″), R^(5″), R^(6″), R^(7″), R^(8″) X″, Z″,n″, o″, p″, r″ and m″ are as defined above—can be prepared by severalknown methods, among them by the method demonstrated on reaction scheme1., by acylation of the compound of formula (III″) by an acylationmethod known from the organic chemistry. As for acylating agentpreferably an acid chloride, for acid binding agent triethyl amineand/or pyridine can be used, but other compounds known as acid binderscan also be applied.

The compound of the general formula (III″)—wherein the meanings ofR^(1″), R^(2″), R^(3″), R^(4″), R^(5″), R^(6″), R^(7″), R^(8″) X″, Z″,n″, o″, p″, r″ and m″ are the same as defined above—can beprepared fromthe compound of formula (IV″), by methods known per se (Nan Zhang,Bioorg. and Med. Chem. Lett., 10, 2825, 2000).

The compound of the general formula (IV″)—wherein the meanings ofR^(4″), R^(5″), R^(6″), R^(7″), R^(8″) X″, Z″, r″, o″, p″ and m″ are asdefined above—can be prepared from the compound of formula (V″), bymethods known per se (D. L. Leysen, J. Heterocyclic Chem., 24, 1611,1987).

The compound of the general formula (V″)—wherein the meanings of R^(4″),R^(5″), R^(7″), R^(8″), Z″, r″, o″, p″ and m″ are as defined above—canbe prepared from the compound of formula (VI″), by methods known per se(Pfizer (Inc) U.S. Pat. No. 4,175,193).

The compound of the general formula (VI″)—wherein the meanings ofR^(4″), R^(5″), R^(7″), R^(8″), Z″, r″, o″, p″ and m″ are as definedabove —can be prepared from the compound of formula (VII″), by methodsknown per se (D. L. Leysen, J. Heterocyclic Chem., 24, 1611, 1987).

The compound of the general formula (VII″)—wherein the meanings ofR^(4″), R^(5″), R^(7″), R^(8″), Z″, r″, o″, p″ and m″ are as definedabove —can be prepared from the compound of formula (VIII″), by methodsknown per se (D. H. Klaubert and J. H. Sellstedt, J. Med. Chem, 24, 742,1981), or are products on the market, such as the compound where R^(4″)and R^(5″) together with the nitrogen atom form a nitro

The compound of the general formula (VIII″)—wherein the meanings ofR^(4″), R^(5″), R^(7″), R^(8″), Z″, r″, o″, p″ and m″ are as definedabove —can be prepared from the compound of formula (IX″), by methodsknown per se (J. H. Hutchinson and J. J. Cook, J. Med. Chem. 39, 4583,1996). The compound of the general formulae (I), (I′) (II″), (III″),(IV″), (V″), (VI″), (VII″), (VIII″), (XIII″) and (XIV″) according to theinvention, as well as their preparation and biological activity aredemonstrated by the examples below, without limiting the claims to theexamples.

Other aspects of the invention include the following intermediates:

Compounds of the general formula (I″), where in the formula the meaningsof R^(1″), R^(2″), R^(3″), R^(6″), R⁷″ R^(8″) X″, n″, m″, o″, p″, and r″are as defined in claim 5. and

R^(4″) stands for hydrogen atom, C₃₋₆ cycloalkyl, benzyl, straight orbranched C₁₋₄ alkyl which is optionally substituted by hydroxy, carboxy,or straight or branched C₁₋₄ alkoxy, amino, or an amino substituted withone or two straight or branched C₁₋₄ alkyl, and R^(5″) stands for aprotective group, or R^(4″) and R^(5″), together with the nitrogen atom,form a group of the general formula a″.), wherein R^(7″) and R^(8″)independently stand for hydrogen atom, straight or branched C₁₋₄ alkylor C₃₋₆ cycloalkyl and Z stands for —NR^(11″), where R^(11″) stands fora protecting group.

Compounds of the general formula (II″), where in the formula themeanings of R^(1″), R^(2″), R^(3″), R^(4″), R^(5″), R^(6″), R⁷″ R^(8″)X″, Z″, n″, m″, o″, p″, and r″ are as defined in claim 5.

Compounds of the general formula (III″), where in the formula themeanings of R^(1″), R^(2″), R^(3″), R^(4″), R^(5″), R^(7″), R^(8″), X″,Z″, n″, m″, o″, p″, and r″ are as defined in claim 5.

Compounds of the general formula (IV″) where in the formula the meaningsof R^(4″), R^(5″), R^(7″), R^(8″), Z″, m″, o″, p″, and r″ are as definedin claim 5.

Compounds of the general formula (V″), where in the formula the meaningsof R^(4″), R^(5″), R^(7″), R^(8″), Z″, m″, o″, p″, and r″ are as definedin claim 5.

Compounds of the general formula (VI″), where in the formula themeanings of R^(4″), R^(5″), R^(7″), R^(8″), Z″, m″, o″, p″, and r″ areas defined in claim 5., with the proviso that R^(4″) and R^(5″) togetherwith the nitrogen atom, have a meaning different from dimethylamino- andamino.

Compounds of the general formula (VII″), where in the formulae themeanings of R^(4″), R^(5″), R^(7″), R^(8″), Z″, m″, o″, p″, and r″ areas defined in claim 5., with the proviso that R^(4″) and R^(5″) togetherwith the nitrogen, atom have a meaning different from dimethylamino-,diethylamino-, and amino.

Compounds of the general formula (VIII″), where in the formula R^(4″)and R^(5″) together with the nitrogen atom, represent a morpholino,N-methylpiperazino, piperazino or 4-benzylpyperazino.

Compounds of the general formula (XIII″), wherein the meanings of thesubstituents R^(1″), R^(2″), R^(3″), R^(6″) and n″ are as defined inclaim 5.

A scheme for preparing compounds according to the invention follows:

EXAMPLES Example 14-Methoxy-N-[6-(morpholin-4-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, group a.) for morpholino, R⁶ for 4-methoxyphenyl, X for NH andthe value of n is 1.

a.) 2-Nitro-5-(morpholin-4-yl)benzoic acid

The mixture of 5 g of 2-nitro-5-chlorobenzoic acid and 15 mL ofmorpholine is stirred at 120° C. for 6 hours. To the reaction mixture150 mL of ethyl acetate is added. The precipitated yellow crystallinematerial is filtered off, dissolved in 15 mL of water. The pH of themixture is adjusted to 6 with acetic acid. The precipitated material isfiltered off, washed with water and dried, to obtain 4.2 g of the titlecompound. Mp.: 172° C.

¹H-NMR (DMSO-d₆) 7.85 ppm (m, 1H), 7.0-6.9 (m, 2H), 3.67 (m, 4H), 2.85(m, 4H).

b.) 2-Amino-5-(morpholin-4-yl)benzoic acid

The mixture of 6 g of 2-nitro-5-(morpholin-4-yl)benzoic acid, 15 mL ofcyclohexene and 3 g of Pd/C (10%) is heated under reflux conditions in120 mL of ethanol for 6 hours. The hot reaction mixture is filteredthrough celite filter. The filtrate is evaporated to obtain 4.8 g of thetitle compound. m.p.: 242° C.

¹H-NMR (DMSO-d₆) 7.25 ppm (m, 1H), 6.96 (m, 1H), 6.62 (m, 1H), 3.69 (m,4H), 2.85 (m, 4H).

c.) 5-(Morpholin-4-yl)isatoic anhydride

To the mixture of 8.9 g of 2-amino-5-(morpholin-4-yl)benzoic acid in 60mL of dioxane, under stirring and external cold water cooling 10 mL ofdiphosgene is added dropwise. The mixture is heated under refluxconditions for 4 hours. From the cold reaction mixture the solidmaterial is filtered off, washed with 50 mL of ether. The product isstirred for 5 minutes in the mixture of 50 mL of methanol and 5 mL oftriethylamine, it is filtered off and washed with 30 mL of methanol.After drying 7 g of the title product is obtained, m.p.: 235° C.

¹H-NMR (DMSO-d₆) 7.8 ppm (m, 1H), 6.78 (m, 1H), 6.66 (m, 1H), 3.73 (m,4H), 2.92 (m, 4H).

d.) 2-Amino-3-cyano-4-hydroxy-6-(morpholin-4-yl)quinoline

4 g of malonitrile is dissolved in 50 mL of dimethylformamide. To thesolution, in several portions, 2.4 g of 60% oily dispersion of sodiumhydride are added. To the clear solution 8 g of5-(morpholin-4-yl)isatoic anhydride is added and the mixture is stirredat room temperature for 10 hours. The reaction mixture is diluted with70 mL of water and extracted with 2×30 mL of ethyl acetate. The aqueousphase is evaporated in vacuum, the solid residue is dissolved in 20 mLof water, the pH is adjusted to 6 with acetic acid. The precipitatedmaterial is filtered off, washed with water. After drying 6.5 g of thetitle compound is obtained, m.p.: 291° C.

¹H-NMR (DMSO-d₆) 7.27 ppm (m, 1H), 7.14 (m, 1H), 7.03 (m, 1H), 3.74 (m,4H), 3.12 (m, 4H).

e.) 2-Amino-3-cyano-4-chloro-6-(morpholin-4-yl)quinoline

The mixture of 1.7 g of2-amino-3-cyano-4-hydroxy-6-(morpholin-4-yl)quinoline and 3.4 mL ofphosphoryl chloride is stirred at 120° C. for 4 hours. The cooledreaction mixture is poured onto 30 g of ice, the pH of the mixture isadjusted to 8 with 10% NaOH solution, and the precipitated material isfiltered off. After drying 1.5 g of the title compound is obtained,m.p.: 206° C.

¹H-NMR (DMSO-d₆) 7.69 ppm (m, 1H), 7.49 (m, 1H), 7.08 (m, 1H), 6.83 (s,2H), 3.74 (m, 4H), 3.08 (M, 4H).

f.) 2-Amino-3-cyano-4-benzylamino-6-(morpholin-4-yl)quinoline

3 g of 2-amino-3-cyano-4-chloro-6-(morpholin-4-yl)quinoline and 6 mL ofbenzylamine are stirred at 125° C. for 3 hours. The reaction mixture ispoured onto 30 mL of water. The precipitated material is filtered off,washed with 20 mL of water. After drying 2.3 g of the title compound isobtained, m.p.: 202° C.

¹H-NMR (DMSO-d₆) 8.14 ppm (m, 1H), 7.5-7.2 (m, 8H), 5.85 (s, 2H), 5.04(d, 2H), 3.65 (m, 4H), 3.1 (m, 4H).

g.)4-Methoxy-N-(4-methoxybenzoyl)-N-(6-(morpholin-4-yl)-4-benzylamino-3-cyanoquinolin-2-yl)benzamide

To the solution of 0.4 g of2-amino-3-cyano-4-benzylamino-6-(morpholin-4-yl)quinoline in 2 mL ofpyridine, 0.4 mL of 4-methoxybenzoyl chloride is added under stirringand cooling. The reaction mixture is stirred at 80° C. for 8 hours, andthen poured onto 5 mL of ice-water. The precipitated material isfiltered off, washed twice with 3 mL of water. After drying 0.53 g ofthe title compound is obtained, m.p.: 157° C.

¹H-NMR (DMSO-d₆) 8.92 ppm (t, 1H), 8.32 (m, 4H), 7.61 (m, 3H), 7.38 (m,5H), 7.12 (m, 4H), 5.1 (d, 2H), 3.82 (m, 8H).

h.)4-Methoxy-N-[(6-(morpholin-4-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

To the solution of 2.3 g of4-methoxy-N-(4-methoxybenzoyl)-N-(6-(morpholin-4-yl)-4-benzylamino-3-cyanoquinolin-2-yl)benzamidein 20 mL of acetonitrile, 5 mL of 1N methanolic potassium hydroxidesolution is added. The reaction mixture is heated under refluxconditions for 10 minutes, 1.5 mL of glacial acetic acid is added to it,then it is neutralized with 15 mL of 1M sodium hydrogen carbonatesolution. The precipitate is filtered off, the yellow crystallinematerial is recrystallized from the mixture of 5 mL of dimethylformamideand 40 mL of ethanol. After drying 1.3 g of the title compound isobtained, m.p.: 260° C.

¹H-NMR (DMSO-d₆) 10.5 ppm (s, 1H), 8.92 (t, 1H), 8.4 (m, 2H), 7.66 (m,3H), 7.35 (m, 5H), 7.05 (, 2H), 5.1 (d, 2H), 3.82 (m, 8H).

Example 24-Methoxy-N-[6-(4-methylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, group a.) for 4-methylpiperazin-1-yl, R⁶ for 4-methoxyphenyl, Xfor NH and the value of N is 1.

a.) 2-Nitro-5-(4-methylpiperazin-1-yl)benzoic acid

The mixture of 10 g of 2-nitro-5-chlorobenzoic acid and 30 mL ofN-methylpiperazine is stirred at 120° C. for 6 hours. To the reactionmixture 150 mL of ethyl acetate is added. The precipitated yellowcrystalline material is filtered off, dissolved in 15 mL of water. ThepH of the mixture is adjusted to 6 with acetic acid. The precipitatedmaterial is filtered off, washed with water and dried, to obtain 11.2 gof the title compound. m.p.: 212° C.

¹H-NMR (DMSO-d₆) 7.89 ppm (d, 1H), 7.03-6.93 (m, 2H), 3.7-3.45 (m, 8H),2.25 (s, 3H).

b.) 2-Amino-5-(4-methylpiperazin-1-yl)benzoic acid

The mixture of 9 g of 2-nitro-5-(4-methylpiperazino)benzoic acid, 20 mLof cyclohexene and 3.5 g of Pd/C (10%) is heated under reflux conditionsin 120 mL of ethanol for 6 hours. The hot reaction mixture is filteredthrough celite filter. The filtrate is evaporated to obtain 3.5 g of thetitle compound. m.p.: 212° C.

¹H-NMR (DMSO-d₆) 7.19 ppm (d, 1H), 7.06 (m, 1H), 6.69 (d, 1H), 2.9 (m,4H), 2.43 (m, 4H), 2.22 (s, 3H).

c.) 5-(4-methylpiperazin-1-yl)isatoic anhydride

To the mixture of 5.3 g of 2-amino-5-(4-methylpiperazino)benzoic acid in30 mL of dioxane, under stirring and external cold water cooling 6 mL ofdiphosgene is added dropwise. The mixture is heated under refluxconditions for 4 hours. From the cold reaction mixture the solidmaterial is filtered off, washed with 50 mL of ether. The product isstirred for 5 minutes in the mixture of 50 mL of methanol and 5 mL oftriethylamine, filtered off, washed with 30 mL of methanol. After drying5.4 g of the title product is obtained, m.p.: 285° C.

¹H-NMR (DMSO-d₆) 7.81 ppm (m, 1H), 6.73 (m, 1H), 6.62 (m, 1H), 3.73-2.92(m, 8H), 2.23 (s, 3H).

d.) 2-Amino-3-cyano-4-hydroxy-6-(4-methylpiperazin-1-yl)quinoline

2 g of malonitrile is dissolved in 30 mL of dimethylformamide. To thesolution, in several portions, 1.3 g of 60% oily dispersion of sodiumhydride are added. To the clear solution 6.5 g of5-(4-methylpiperazino)isatoic acid anhydride is added and the mixture isstirred at room temperature for 10 hours. The reaction mixture isdiluted with 70 mL of water and extracted with 2×30 mL of ethyl acetate.The aqueous phase is evaporated in vacuum, the solid residue isdissolved in 20 mL of water, the pH is adjusted to 6 with acetic acid.The precipitated material is filtered off, washed with water. Afterdrying 5.2 g of the title compound is obtained, m.p.: 156° C.

hu 1H-NMR (DMSO-d₆) 7.23 ppm (m, 1H), 7.12 (m, 1H), 7.03 (m, 1H),3.65-2.83 (m, 8), 2.1 (s, 3H).

e.) 2-Amino-3-cyano-4-chloro-6-(4-methylpiperazin-1-yl)quinoline

The mixture of 2 g of2-amino-3-cyano-4-hydroxy-6-(4-methylpiperazino)quinoline and 4 mL ofphosphoryl chloride is stirred at 120° C. for 4 hours. The cooledreaction mixture is poured onto 40 g of ice, the pH of the mixture isadjusted to 8 with 10% NaOH solution, and the precipitated material isfiltered off. After drying 1.5 g of the title compound is obtained,m.p.: 189° C.

¹H-NMR (DMSO-d₆) 7.69 ppm (m, 1H), 7.49 (m, 1H), 7.08 (m, 1H), 6.83 (s,2H), 3.25-2.57 (m, 8H), 2.29 (s, 3H).

f.) 2-Amino-3-cyano-4-benzylamino-6-(4-methylpiperazin-1-yl)quinoline

3 g of 2-amino-3-cyano-4-chloro-6-(4-methylpiperazino)quinoline and 6 mLof benzylamine are stirred at 125° C. for 3 hours. The reaction mixtureis poured onto 30 mL of water. The precipitated material is filteredoff, washed with 20 mL of water. After drying 2.3 g of the titlecompound is obtained, m.p.: 176° C.

¹H-NMR (DMSO-d₆) 8.5 ppm (t, 1H), 7.5-7.15 (m, 8H), 5.85 (s, 2H), 5.04(d, 2H), 3.65-3.12 (m, 8H), 2.23 (s, 3H).

g.)4-Methoxy-N-(4-methoxybenzoyl)-N-[6-(4-methylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

To the solution of 0.6 g of2-amino-3-cyano-4-benzylamino-6-(4-methylpiperazino)quinoline in 2 mL ofpyridine, 0.6 mL of 4-methoxybenzoyl chloride is added under stirringand cooling. The reaction mixture is stirred at 80° C. for 8 hours, andthen poured onto 5 mL of ice-water. The precipitated material isfiltered off, washed twice with 3 mL of water. After drying 0.63 g ofthe title compound is obtained, m.p.: 176° C.

¹H-NMR (DMSO-d₆) 8.39 ppm (m, 1H), 7.95 (m, 2H), 7.47 (m, 5H), 7.32 (m,5H), 7.14 (m, 4H), 5.1 (m, 2H), 3.82 (s, 3H), 3.52-2.98 (m, 8H), 2.25(s, 3H).

h.)4-Methoxy-N-[6-(4-methylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

To the solution of 2.3 g of4-methoxy-N-(4-methoxybenzoyl)-N-(6-(4-methylpiperazino)-4-benzylamino-3-cyanoquinolin-2-yl)benzamidein 15 mL of acetonitrile, 4 mL of 1N methanolic potassium hydroxidesolution is added. The reaction mixture is heated under refluxconditions for 10 minutes, 1 mL of glacial acetic acid is added to it,then it is neutralized with 12 mL of 1M sodium hydrogen carbonatesolution. The precipitate is filtered off, the yellow crystallinematerial is recrystallized from the mixture of 15 mL of methanol and 35mL of water. After drying 1.1 g of the title compound is obtained, m.p.:173° C.

¹H-NMR (DMSO-d₆) 10.53 ppm (m, 1H), 8.39 (m, 1H), 7.95 (m, 2H), 7.57 (m,3H), 7.34 (m, 5H), 7.04 (m, 2H), 5.1 (m, 2H), 3.82 (s, 3H), 3.52-2.98(m, 8H), 2.25 (s, 3H).

Example 34-Methoxy-N-(6-dimethylamino-4-benzylamino-3-cyanoquinolin-2-yl)benzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, R⁴ and R⁵ for methyl group, R⁶ for 4-methoxyphenyl, X for NH andthe value of n is 1.

a.) 2-Nitro-5-dimethylaminobenzoic acid

The mixture of 5 g of 2-nitro-5-chlorobenzoic acid and 15 mL of 60%aqueous dimethylamine solution is stirred at 100° C. for 6 hours. Thereaction mixture is evaporated, the residue dissolved in 15 mL of water.The pH of the mixture is adjusted to 6 with acetic acid. Theprecipitated yellow crystalline material is filtered off, washed withwater and dried, to obtain 3.4 g of the title compound. Mp.: 189° C.

¹H-NMR (DMSO-d₆) 7.78 ppm (d, 1H), 6.59 (m, 1H), 6.48 (m, 1H), 3.0 (s,6H).

b.) 2-Amino-5-dimethylaminobenzoic acid

The mixture of 2.1 g of 2-nitro-5-dimethylaminobenzoic acid, 7 mL ofcyclohexene and 1.5 g of Pd/C (10%) is heated under reflux conditions in60 mL of ethanol for 6 hours. The hot reaction mixture is filteredthrough celite filter. The filtrate is evaporated to obtain 1.1 g of thetitle compound. M.p.: 232° C.

¹H-NMR (DMSO-d₆) 7.01 ppm (m, 1H), 6.84 (m, 1H), 6.78 (m, 1H), 2.88 (s,6H).

c.) 5-Dimethylaminoisatoic anhydride

To the mixture of 8.9 g of 2-amino-5-dimethylaminobenzoic acid in 60 mLof dioxane, under stirring and external cold water cooling 10 mL ofdiphosgene is added dropwise. The mixture is heated under refluxconditions for 4 hours. From the cold reaction mixture the solidmaterial is filtered off, washed with 50 mL of ether. The product isstirred for 5 minutes in the mixture of 50 mL of methanol and 5 mL oftriethylamine, filtered off, washed with 30 mL of methanol. After drying7 g of the title product is obtained, m.p.: 258° C.

¹H-NMR (DMSO-d₆) 7.56 ppm (m, 1H), 7.42 (m, 1H), 7.13 (m, 1H), 2.97 (s,6H).

d.) 2-Amino-3-cyano-4-hydroxy-6-dimethylaminoquinoline

To the solution of 4 g of malonitrile in 50 mL of dimethylformamide inseveral portions 2.4 g of sodium hydride 60% oily dispersion is added.To the clear solution 8 g of 5-dimethylamino-isatoic anhydride is addedand the mixture is stirred at room temperature for 10 hours. The mixtureis diluted with 70 mL of water and extracted with 2×30 mL of ethylacetate. The aqueous phase is evaporated in vacuum, the solid residue isdissolved in 20 mL of water, the pH is adjusted to 6 with acetic acid.The precipitated material is filtered off, and washed with water. Afterdrying 6.5 g of the title compound is obtained, m.p.: 360° C.

¹H-NMR (DMSO-d₆) 7.43 ppm(m, 1H), 7.23(m, 1H), 7.11(m, 1H), 2.95(s, 6H).

e.) 2-Amino-3-cyano-4-chloro-6-dimethylaminoquinoline

The mixture of 1.7 g of2-amino-3-cyano-4-hydroxy-6-dimethylaminoquinoline and 3.4 mL ofphosphoryl chloride is stirred at 120° C. for 4 hours. The cooledreaction mixture is poured onto 30 g of ice, the pH of the mixture isadjusted to 8 with 10% NaOH solution and the precipitated material isfiltered off. After drying 1.5 g of the title compound is obtained,m.p.: 285° C.

¹H-NMR (DMSO-d₆) 7.43 ppm(mm, 1H), 7.21(m, 1H), 7.05(m, 1H), 6.75(s,2H), 2.99(s, 6H).

f.) 2-Amino-3-cyano-4-benzylamino-6-dimethylaminoquinoline

3 g of 2-amino-3-cyano-4-chloro-6-dimethylaminoquinoline and 6 mL ofbenzylamine are stirred at 125° C. for 3 hours. The reaction mixture ispoured onto 30 mL of water. The precipitated material is filtered off,washed with 20 mL of water. After drying 2.3 g of the title compound isobtained, m.p.: 265° C.

¹H-NMR (DMSO-d₆) 8.55-8.45 ppm(m, 2H), 7.8(m, 1H), 7.5-7.23(m, 7H),6.25(s, 2H), 5.08(d, 2H), 2.99(s, 6H).

g.)4-Methoxy-N-(4-methoxybenzoyl)-N-(6-dimethylamino-4-benzylamino-3-cyanoquinolin-2-yl)benzamide

To the solution of 0.4 g of2-amino-3-cyano-4-benzylamino-6-dimethylaminoquinoline in 2 mL ofpyridine, 0.4 mL of 4-methoxybenzoyl chloride is added under stirringand cooling. The reaction mixture is stirred at 80° C. for 8 hours, thenpoured onto 5 mL of ice-water. The precipitated material is filteredoff, washed twice with 3 mL of water. After drying 0.53 g of the titlecompound is obtained, m.p.: 156° C.

¹H-NMR (DMSO-d₆) 8.35 ppm(m, 1H), 7.9(m, 2H), 7.47(m, 5H), 7.3(m, 5H),7.1(m, 4H), 5.12(m, 2H), 3.82(s, 3H), 3.0(s, 6H).

h.)4-Methoxy-N-(6-dimethylamino-4-benzylamino-3-cyanoquinolin-2-yl)benzamide

To the solution of 2.3 g of4-methoxy-N-(4-methoxybenzoyl)-N-(6-dimethylamino-4-benzylamino-3-cyanoquinolin-2-yl)benzamidein 20 mL of acetonitrile, 5 mL of 1N methanolic potassium hydroxidesolution is added. The reaction mixture is heated under refluxconditions for 10 minutes, 1.5 mL of glacial acetic acid is added to it,then it is neutralized with 15 mL of 1M sodium hydrogen carbonatesolution. The precipitate is filtered off, the yellow crystallinematerial is recrystallized from the mixture of 5 ml of dimethylformamideand 40 mL of ethanol. After drying 1.3 g of the title compound isobtained, m.p.: 185° C.

¹H-NMR (DMSO-d₆) 10.5 ppm(m, 1H), 8.35(m, 1H), 7.91(m, 2H), 7.53(m, 3H),7.3(m, 5H), 7.02(m, 2H), 5. 1(m, 2H), 3.85(s, 3H), 3.0(s, 6H).

Example 44-Methoxy-N-(6-dimethylamino-4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)benzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ for2-furyl group, R⁴ and R⁵ for methyl group, R⁶ for 4-methoxyphenyl, X forNH and the value of n is 1.

a.) 2-Amino-3-cyano-4-[2-furylmethylamino]-6-dimethylaminoquinoline

3 g of 2-amino-3-cyano-4-chloro-6-dimethylaminoquinoline and 6 mL offurfurylamine are stirred at 125° C. for 3 hours. The reaction mixtureis poured onto 30 mL of water. The precipitated material is filteredoff, washed with 20 mL of water. After drying 2.05 g of the titlecompound is obtained, m.p.: 235° C.

¹H-NMR (DMSO-d₆) 8.7 ppm(m, 1H), 7.6(m, 1H), 7.35-7.23(m, 3H), 6.8 (s,2H), 6.4(m, 2H), 5.06(d, 2H), 2.96(s, 6H).

b.)4-Methoxy-N-(4-methoxybenzoyl)-N-(6-dimethylamino-4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)benzamide

To the solution of 0.4 g of2-amino-3-cyano-4-[2-furylmethylamino]-6-dimethylaminoquinoline in 2 mLof pyridine, 0.4 mL of 4-methoxybenzoyl chloride is added under stirringand cooling. The reaction mixture is stirred at 80° C. for 8 hours, thenpoured onto 5 mL of ice-water. The precipitated material is filteredoff, washed twice with 3 mL of water. After drying 0.5 g of the titlecompound is obtained, m.p.: 143° C.

¹H-NMR (DMSO-d₆) 8.35 ppm(m, 1H), 7.9-7.1(m, 14H), 5.12(m, 2H), 3.82(s,6H), 3.0(s, 6H).

c.)4-Methoxy-N-(6-dimethylamino-4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)benzamide

To the solution of 2.3 g of4-methoxy-N-(4-methoxybenzoyl)-N-(6-dimethylamino-4-[2-furylmethylamino]-3-cyanoquinolin-2-yl)benzamidein 20 mL of acetonitrile, 5 mL of 1N methanolic potassium hydroxidesolution is added. The reaction mixture is heated under refluxconditions for 10 minutes, 1.5 mL of glacial acetic acid is added to it,then it is neutralized with 15 mL of 1M sodium hydrogen carbonatesolution. The precipitate is filtered off, the yellow crystallinematerial is recrystallized from the mixture of 5 ml of dimethylformamideand 40 mL of ethanol. After drying 1.1 g of the title compound isobtained, m.p.: 195° C.

¹H-NMR (DMSO-d₆) 10.5 ppm(m, 1H), 8.25(t, 1H), 7.98(m, 2H), 7.63-7.03(m,7H), 6.42 (d, 1H), 5.04(d, 2H), 3.85(s, 3H), 3.05(s, 6H).

Example 54-Methoxy-N-(6-dimethylamino-4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ for2-thienyl group, R⁴ and R⁵ for methyl group, R⁶ for 4-methoxyphenyl,Xfor NH and the value of n is 1.

a.)-2-Amino-3-cyano-4-[2-thienylmethylamino]-6-dimethylaminoquinoline

3 g of 2-amino-3-cyano-4-chloro-6-dimethylaminoquinoline and 6 mL of2-thienylmethylamine are stirred at 125° C. for 3 hours. The reactionmixture is poured onto 30 mL of water. The precipitated material isfiltered off, washed with 20 mL of water. After drying 1.9 g of thetitle compound is obtained, m.p.: 211° C.

¹H-NMR (DMSO-d₆) 8.2 ppm(m, 1H), 7.46-6.95(m, 6H), 6.08(s, 2H), 5.18(d,2H), 2.94(s, 6H).

b.)4-Methoxy-N-(4-methoxybenzoyl)-N-(6-dimethylamino-4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamide

To the solution of 0.4 g of2-amino-3-cyano-4-[2-thienylmethylamino]-6-dimethylaminoquinoline in 2mL of pyridine, 0.4 mL of 4-methoxybenzoyl chloride is added understirring and cooling. The reaction mixture is stirred at 80° C. for 8hours, then poured onto 5 mL of ice-water. The precipitated material isfiltered off, washed twice with 3 mL of water. After drying 0.43 g ofthe title compound is obtained, m.p.: 171° C.

¹H-NMR (DMSO-d₆) 8.35 ppm(m, 1H), 7.9-7.05(m, 14H), 5.12(m, 2H), 3.82(s,6H), 3.0(s, 6H).

c.)4-Methoxy-N-(6-dimethylamino-4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamide

To the solution of 2.3 g of4-methoxy-N-(4-methoxybenzoyl)-N-(6-dimethylamino-4-[2-thienylmethylamino]-3-cyanoquinolin-2-yl)benzamidein 20 mL of acetonitrile, 5 ML of 1N methanolic potassium hydroxidesolution is added. The reaction mixture is heated under refluxconditions for 10 minutes, 1.5 mL of glacial acetic acid is added to it,then it is neutralized with 15 mL of 1M sodium hydrogen carbonatesolution. The precipitate is filtered off, the yellow crystallinematerial is recrystallized from the mixture of 5 ml of dimethylformamideand 40 mL of ethanol. After drying 1.15 g of the title compound isobtained, m.p.: 163° C.

¹H-NMR (DMSO-d₆) 10.5 ppm(m, 1H), 8.3(t, 1H), 7.98(m, 2H), 7.63-6.96(m,8H), 5.2(d, 2H), 3.85(s, 3H), 3.05(s, 6H).

Example 64-Methoxy-N-[6-(piperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, group a.) for piperazino, R⁶ for 4-methoxyphenyl, X for NH andthe value of n is 1.

a.) 2-Nitro-5-(4-benzylpiperazin-1-yl)benzoic acid

The mixture of 20 g of 2-nitro-5-chlorobenzoic acid and 50 mL ofN-benzylpiperazine is stirred at 120° C. for 6 hours. To the reactionmixture 250 mL of ethyl acetate is added. The precipitated yellowcrystalline material is filtered off, dissolved in 200 mL of water. ThepH of the mixture is adjusted to 6 with acetic acid. The precipitatedmaterial is filtered off, washed with water and dried, to obtain 30 g ofthe title compound. M.p.: 172° C.

¹H-NMR (DMSO-d₆) 7.8-6.7 ppm (m, 8H), 3.5 (s, 2H), 3.5-2.8 (m, 8H).

b.) 2-Amino-5-(4-benzylpiperazin-1-yl)benzoic acid

The mixture of 6 g of 2-nitro-5-(4-benzylpiperazin-1-yl)benzoic acid, 15mL of cyclohexene and 3 g of Pd/C (10%) is heated under refluxconditions in 120 mL of ethanol for 6 hours. The hot reaction mixture isfiltered through celite filter. The filtrate is evaporated to obtain 4.8g of the title compound. m.p.: 242° C.

¹H-NMR (DMSO-d₆) 7.5-6.8 ppm (m, 8H), 3.68 (s, 3H), 3.5-2.95 (m, 8H).

c.) 5-(4-benzylpiperazin-1-yl)isatoic anhydride

To the mixture of 15 g of 2-amino-5-(4-benzylpiperazin-1-yl)benzoic acidin 90 mL of dioxane, under stirring and external cold water cooling 12.7mL of diphosgene is added dropwise. The mixture is heated under refluxconditions for 4 hours. From the cold reaction mixture the solidmaterial is filtered off, washed with 120 mL of ether. The product isstirred for 5 minutes in the mixture of 100 mL of methanol and 10 mL oftriethylamine, it is filtered off and washed with 50 mL of methanol.After drying 17 g of the title product is obtained, m.p.: 235° C.

¹H-NMR (DMSO-d₆) 7.68-7.1 ppm (m, 8H), 3.6 (s, 2H), 3.5-2.5(m, 8H).

d.) 2-Amino-3-cyano-4-hydroxy-6-(4-benzylpiperazin-1-yl)quinoline

6.1 g of malonitrile is dissolved in 100 mL of dimethylformamide. To thesolution, in several portions, 3.6 g of 60% oily dispersion of sodiumhydride are added. To the clear solution 18 g of5-(4-benzylpiperazin-1-yl)isatoic anhydride is added and the mixture isstirred at room temperature for 10 hours. The reaction mixture isdiluted with 100 mL of water and extracted with 2×50 mL of ethylacetate. The aqueous phase is evaporated in vacuum, the solid residue isdissolved in 50 mL of water, the pH is adjusted to 6 with acetic acidand refluxed for 5 hours. After cooling the precipitated material isfiltered off, washed with water. After drying 14.3 g of the titlecompound is obtained, m.p.: 291° C.

¹H-NMR (DMSO-d₆) 7.7-6.5 ppm (m, 10H), 4.2 (s, 2H), 3.5-2.5 (m, 8H),

e.) 2-Amino-3-cyano-4-chloro-6-(4-benzylpiperazin-1-yl)quinoline

The mixture of 14 g of2-amino-3-cyano-4-hydroxy-6-(4-benzylpiperazin-1-yl)quinoline and 28 mLof phosphoryl chloride is stirred at 120° C. for 6 hours. The cooledreaction mixture is poured onto 500 g of ice, the pH of the mixture isadjusted to 8 with 10% NaOH solution, and the precipitated material isfiltered off. After drying 14.5 g of the title compound is obtained,m.p.: 206° C.

¹H-NMR (DMSO-d₆) 7.7-7.14 ppm (m, 8H), 6.9 (s, 2H), 3.7 (s, 2H), 3.5-2.5(m, 8H).

f.) 2-Amino-3-cyano-4-benzylamino-6-(4-benzylpiperazin-1-yl)quinoline

14 g of 2-amino-3-cyano-4-chloro-6-(4-benzylpiperazin-1-yl)quinoline and28 mL of benzylamine are stirred at 125° C. for 4 hours. The reactionmixture is poured onto 100 mL of water. The precipitated material isfiltered off, washed with 2×50 mL of water. After drying 8 g of thetitle compound is obtained, m.p.: 202° C.

¹H-NMR (DMSO-d₆) 8.1 ppm (m, 1H), 7.5-7.2 (m, 13H), 5.8 (s, 2H), 5.0 (d,2H), 3.54 (s,2H), 3.5-2.5 (m, 8H).

g.)4-Methoxy-N-(4-methoxybenzoyl)-N-(6-(4-benzylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl)benzamide

To the solution of 0.9 g of2-amino-3-cyano-4-benzylamino-6-(4-benzylpiperazin-1-yl)quinoline in 5mL of ethylacetate containing 0.3 mL of triethylamine, 0.5 mL of4-methoxybenzoyl chloride is added under stirring and cooling. Thereaction mixture is stirred at 80° C. for 8 hours, then poured onto 10mL of ice-water. The precipitated material is filtered off, washed twicewith 3 mL of water. After drying 0.53 g of the title compound isobtained, m.p.: 157° C.

¹H-NMR (DMSO-d₆) 8.92 ppm (t, 1H), 8.32 (m, 4H), 7.61 (m, 3H), 7.38 (m,5H),7.12 (m, 4H), 5.1 (d, 2H), 3.82 (s, 6H), 3.56 (s, 2H), 3.5-2.5 (m,8H).

h.)4-Methoxy-N-[(6-(4-benzylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

To the solution of 2.0 g of4-methoxy-N-(4-methoxybenzoyl)-N-(6-(4-benzylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl)benzamidein 20 mL of acetonitrile, 4 mL of 1N methanolic potassium hydroxidesolution is added. The reaction mixture is heated under refluxconditions for 10 minutes, 1.5 mL of glacial acetic acid is added to it,then it is neutralized with 15 mL of 1M sodium hydrogen carbonatesolution. The precipitate is filtered off, the yellow crystallinematerial is recrystallized from the mixture of 5 mL of dimethylformamideand 40 mL of ethanol. After drying 1.3 g of the title compound isobtained, m.p.: 260° C.

¹H-NMR (DMSO-d₆) 10.5 ppm (s, 1H), 8.92 (t, 1H), 7.97-7.01 (m, 17H), 5.1(d, 2H), 3.82 (s, 3H), 3.56 (s, 2H), 3.5-2.5 (m, 8H).

i.)4-Methoxy-N-[(6-(piperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

1 g of4-methoxy-N-[(6-(4-benzylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamidewas dissolved in dimethylformamide and water, containing 0.5 mL ofacetic acid and was hydrogenated in the presence of 50 mg of Pd/C (10%)for 4 hours at 45° C. After filtration of the catalyst and evaporationof the solvent the crude product was purified by chromathography to give0.65 g of the title compound, mp: 145° C.

¹H-NMR (DMSO-d₆) 10.5 ppm (s, 1H), 8.48 (m, 1H), 7.94 (m, 2H), 7.6-7.35(m, 2H), 7.33 (s, 5H), 7.24 (m, 1H), 7.02 (m, 2H), 5.08 (d, 2H), 3.83(s, 3H), 3.22 (m, 4H), 2.86 (m, 4H).

Example 7N-[6-amino-4-(benzylamino)-3-cyanoquinolin-2-yl]-4-methoxybenzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, NR⁴R⁵ stands for NH₂, R⁶ for 4-methoxyphenyl, X for NH and thevalue of n is 1.

a,) 5-Nitroisatoic anhydride

To the mixture of 37 g of 2-amino-5-nitrobenzoic acid in 500 mL ofdioxane, under stirring 24.5 mL of diphosgene was added dropwise. Themixture was heated under reflux conditions for 6 hours then it wasevaporated to dryness. The residue was suspended in 100 mL of diethylether and filtered off to give 41.4 g of the title product as HCl salt,m.p.: 256-259° C. LC-MS: MH⁺ 209; Ret. time: 5 minutes.

¹H-NMR (DMSO-d₆) 8.54 (d, 1H), 8.46 (dd, 1H), 7.37 (d, 1H) ppm.

b,) 2-Amino-3-cyano-4-hydroxy-6-nitroquinoline

To the solution of 47.25 g of 5-nitroisatoic anhydride in 230 mL of DMF15 g of malonitrile and 63.3 mL of triethylamine were added and thereaction mixture was stirred at 60° C. for 2 hours. The solvent wasevaporated at reduced pressure and the residue was mixed with 570 mL ofacetonitrile and 114 mL of concd. HCl and the solution was stirred atroom temperature overnight. The precipitate was then filtered off,washed with water and ethanol to give 49.7 g of product, m.p.: >360° C.LC-MS: MH⁺ 231; Ret. time: 4.78 min.

¹H-NMR (DMSO-d₆) 8.64 (d, 1H), 8.36 (dd, 1H), 7.6 (s, 2H), 7.55 (d, 1H)ppm.

c,) 2-Amino-3-cyano-4-chloro-6-nitroquinoline

The stirred mixture of 48.5 g of2-amino-3-cyano-4-hydroxy-6-nitroquinoline and 550 mL of phosphorylchloride was refluxed for 4 hours. The reaction mixture was concentratedto half of its volume and the residue was poured onto 1500 g of ice. Theyellow precipitate was filtered off, washed with water to give 60.5 g ofsolid. 25 g of this solid was stirred at room temperature in 500 mL of0.5 N HCl for 6 hours, then the precipitate was filtered off, washedwith water to give 18.5 g of product, m.p.: >360° C. LC-MS: MH⁺ 249;Ret. time: 5.94 min.

¹H-NMR (DMSO-d₆) 8.73 (d, 1H), 8.48 (dd, 1H), 7.92 (d, 1H) ppm.

d,) 2-Amino-3-cyano-4-benzylamino-6-nitroquinoline

A mixture of 7 g of 2-amino-3-cyano-4-chloro-6-nitroquinoline and 30 mLof benzylamine was stirred at 55° C. for 1.5 hours. The reaction mixtureis poured onto 30 mL of water. The precipitated material is filteredoff, washed with water to give 7 g of the title compound, m.p.: 280-283°C. LC-MS: MH⁺ 2320; Ret. time: 5.19 minutes.

¹H-NMR (DMSO-d₆): 9.28 (d, 1H), 8.90 (t, 1H), 8.24 (dd, 1H), 7.3 (m,5H), 6.98 (s,2H), 5.04 (d, 2H) ppm.

e,)4-Methoxy-N-(4-methoxybenzoyl)-N-(4-benzylamino-3-cyano-6-nitroquinolin-2-yl)benzamide

A suspension of 3.5 g of 2-amino-3-cyano-4-benzylamino-6-nitroquinolinein 50 mL of dry pyridine was refluxed with 5.6 g of 4-methoxybenzoylchloride for 3.5 hours. The solvent was evaporated in reduced pressureand the residue was suspended in 30 mL of a saturated solution of Na₂CO₃in water. The suspension was extracted with 3×25 mL of CH₂Cl₂. Thecombined organic extract was evaporated to dryness, the residue wasdissolved in diethyl ether and the solution was kept in a refrigeratorfor a night. The precipitate was filtered off, washed with diethyl etherto give 6.25 g of the title compound, m.p.: 145-148° C. LC-MS: MH⁺ 588;Ret. time: 7.00 min.

f,) N-[(4-Benzylamino-3-cyano-6-nitroquinolin-2-yl]-4-methoxy-benzamide

A suspension of 6.2 g ofN-(4-methoxybenzoyl)-N-(4-benzylamino-3-cyano-6-nitroquinolin-2-yl)-4-methoxybenzamidein 75 mL of acetonitrile was refluxed with 25.2 mL of 1N methanolicpotassium hydroxide solution for 6 minutes. 4.4 mL of glacial aceticacid was dropped to the solution while hot, cooled and neutralized with56.3 mL of 1M aqueous NaHCO₃ solution. The precipitate was filtered off,washed with water to give 4.1 g of title compound, m.p.: 264-266° C.

¹H-NMR (DMSO-d₆) 10.9 (s, 1H), 9.55 (d, 1H), 9.28 (t, 1H), 8.77 (d, 1H),8.48 (dd, 1H), 8.00 (d, 2H), 7.90 (d, 1H), 7.4 (m, 4H), 7.29 (m, 1H),7.05 (d, 2H), 5.12 (d, 2H), 3.84 (s, 3H) ppm.

g,) N-[6-amino-4-(benzylamino)-3-cyanoquinolin-2-yl]-4-methoxybenzamide

A mixture of 320 mL of ethanol, 80 mL of water, 200 mL of THF, 7.6 g ofFe-powder, and 1 mL of concd. HCl was refluxed for 10 min. 9 g ofN-[(4-benzylamino-3-cyano-6-nitroquinolin-2-yl]-4-methoxy-benzamide wasthen added and the reaction mixture was intensively refluxed for 2 hoursand it was cooled to 35° C. The precipitate was filtered off, washedwith a 1:1 mixture of CH₂Cl₂ and ethanol. The combined organic solutionwas filtered on charcoal and evaporated to dryness. The residue wassuspended in a mixture of 50 mL of water and 10 mL of ethanol, the solidmaterial was filtered off, washed with water to give 6.65 g of the titlecompound, mp: 228-230° C. LC-MS: MH⁺ 424; Ret. time: 5.33 minutes.

¹H-NMR (DMSO-d₆) 10.44 (s, 1H), 8.0 (s+d, 3H), 7.50 (d, 1H), 7.3-7.1 (m,7H), 7.03 (d, 2H), 5.55 (s, 2H), 5.04 (d, 2H), 3.83 (s, 3H) ppm.

Example 8N-{6-amino-4-[(2-thienylmethyl)amino)]-3-cyanoquinolin-2-yl}-4-methoxybenzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ for2-thienyl, NR⁴R⁵ stands for NH₂, R⁶ for 4-methoxyphenyl, X for NH andthe value of n is 1.

a,) 2-Amino-3-cyano-4-(2-thienylmethyl)amino-6-nitroquinoline

Applying the same procedure as given under Example 7d with themodification that instead of benzylamine, (2-thienylmethyl)amine wasused to give 7.1 g of the title compound, m.p.: 277-280° C. LC-MS: MH⁺326; Ret. time: 5.25 min.

b,)N-(4-Methoxybenzoyl)-N-[3-cyano-6-nitro-4-(2-thienylmethyl)-aminoquinolin-2-yl]-4-methoxybenzamide

Started the reaction from2-amino-3-cyano-4-(2-thienylmethyl)amino-6-nitroquinoline (3.6 g) andusing the procedure as given for Example 7e, 6.3 g of the title compoundwas prepared, m.p.: 174-177° C. LC-MS: MH⁺ 595; Ret. time: 7.16 min.

c,)N-[3-Cyano-6-nitro-4-(2-thienylmethyl)aminoquinolin-2-yl]-4-methoxybenzamide

Started the reaction from N-(4-methoxybenzoyl)-N-[3-cyano-6-nitro4-(2-thienylmethyl)amino]-quinolin-2-yl)-4-methoxybenzamide (6.2 g) andusing the procedure as given for Example 7f, 4.3 g of the title compoundwas prepared, m.p.: 217-220° C. LC-MS: MH⁺ 460; Ret. time: 6.57 minutes.

¹H-NMR (DMSO-d₆): 11.0 (s, 1H), 9.50 (s, 1H), 9.29 (s, 1H), 8.48 (dd,1H), 8.02 (d, 2H), 7.90 (d, 1H), 7.48 (dd, 1H), 7.21 (d, 1H), 7.0 (m,3H), 5.27 (s, 2H), 3.85 (s, 3H) ppm

d,)N-{6-amino-4-[(2-thienylmethyl)amino]-3-cyanoquinolin-2-yl}-4-methoxybenzamide

Started the reaction fromN-[4-(2-thienylmethyl)amino-3-cyano-6-nitroquinolin-2-yl]-4-methoxybenzamide(10 g) and using the procedure as given for Example 7g, 9.3 g of thetitle compound was prepared, mp: 200-203° C. LC-MS: MH⁺ 430; Ret. time:5.52 minutes.

¹H-NMR (DMSO-d₆) 10.48 (s, 1H), 7.98 (s+d, 3H), 7.50 (d, 1H), 7.41 (dd,1H), 7.21 (s, 1H), 7.16 (dd, 1H), 7.11 (d, 1H), 7.04 (d, 2H), 6.98 (dd,1H), 5.45 (s, 2H), 5.18 (d, 2H), 3.84 (s, 3H) ppm

Example 9N-{4-(benzylamino)-3-cyano-6-(methylamino)quinolin-2-yl}-4-methoxybenzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, R⁴ stands for Me, R⁵ stands for H and R⁶ for 4-methoxyphenyl, Xfor NH and the value of n is 1.

A suspension of 1 g ofN-{4-(benzylamino)-3-cyano-6-aminoquinolin-2-yl}-4-methoxybenzamide and0.65 g of paraformaldehyde in a mixture of 48 mL of ethanol and 48 mL ofCH₂Cl₂ was refluxed with 0.5 mL of 54% HBF₄ solution in diethyl etherfor 1.5 hours. After addition of 0.3 g of NaBH₄ the reflux was continuedfor 1.5 hours followed by addition of a further amount of 0.3 g of NaBH₄and reflux for 2 hours. After repeated addition of 0.25 mL of 54% HBF₄solution in diethyl ether and 0.3 g of NaBH₄ the reflux was continuedfor additional 1.5 hours. The reaction mixture was filtered while hot,the solid was washed with a 1:1 mixture of CH₂Cl₂ and ethanol. Thecombined organic solution was concentrated and the residue waschromatographed on silica gel with a 50:1 mixture of CHCl₃ and ethylacetate to give the crude product. It was recrystallized from a 2:1mixture of ethanol and DMF to give 0.6 g of title compound, m.p.:237-240° C. LC-MS: MH⁺ 438; Ret. time: 5.76 minutes.

¹H-NMR (DMSO-d₆) 10.43 (s, 1H), 8.18 (t, 1H), 7.94 (d, 2H), 7.50 (d,1H), 7.3-7.0 (m, 9H), 6.24 (m, 1H), 5.08 (d, 2H), 3.83 (s, 3H), 2.83 (d,3H) ppm.

Example 10N-{3-Cyano-6-(formylamino)-4-[(2-thienylmethyl)amino]quinolin-2-yl]-4-methoxybenzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ for2-thienyl, R⁴ stands for hydrogen, R⁵ stands for CHO and R⁶ for4-methoxyphenyl, X for NH and the value of n is 1.

A suspension of 0.32 g ofN-{6-amino-4-[(2-thienylmethyl)amino]-3-cyanoquinolin-2-yl}-4-methoxybenzamidewas kept in 60 mL of methyl formate at 100° C. in a closed apparatus(internal pressure: 10 bar) for 7 hours. After cooling the solvent wasevaporated and the residue was recrystallized from CH₂Cl₂/MeOH to give0.22 g of title compound, m.p.: 223-226° C. LC-MS: MH⁺ 458; Ret. time:5.71 min.

Example 11{4-(Benzylamino)-3-cyano-2-[(4-methoxybenzoyl)amino]quinolin-6-yl}sulfamicacid

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, R⁴ stands for H, R⁵ stands for SO₂OH and R⁶ for 4-methoxyphenyl,X for NH and the value of n is 1.

To a solution of 5.2 g of Na₂S₂O₄ (purity: 85%) in 160 mL of water and200 mL of ethanol was added 20 mL of aqueous 1N NaOH solution. Thesolution was heat up to boiling then 2.27 g ofN-[(4-benzylamino-3-cyano-6-nitroquinolin-2-yl]-4-methoxybenzamide wasadded and refluxing was continued for 2 hours. The reaction mixture letstand at room temperature overnight then it was concentrated to half ofits volume. The residue was made alkaline (pH=8) by addition of 1N NaOHsolution and was extracted with CH₂Cl₂. The aqueous solution wasacidified to pH=5 by addition of concd. HCl solution and the precipitatewas filtered off. The solid was chromatographed on silica gel elutedfirst by a mixture of ethyl acetate/methanol/25% ammonia=220/30/2, then200/80/2 to get 0.5 g of title compound as ammonium salt, m.p.: 220-222°C. LC-MS: MH⁺ 504; Ret. time: 5.56 min.

¹H-NMR (DMSO-d₆) 10.48 (s, 1H), 8.21 (s, 1H), 8.09 (t, 1H), 7.95 (d,2H), 7.70 (s, 1H), 7.62 (d, 1H), 7.56 (d, 1H), 7.3 (m, 4H), 7.25 (m,1H), 7.03 (d, 2H), 5.05 (d, 2H), 3.83 (s, 3H) ppm

Example 12{4-(Benzylamino)-3-cyano-2-[(4-methoxybenzoyl)amino]quinolin-6-yl}methylsulfamicacid

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, R⁴ stands for Me, R⁵ stands for SO₂OH and R⁶ for4-methoxyphenyl, X for NH and the value of n is 1.

A solution of 110 mg ofN-{4-(benzylamino)-3-cyano-6-(methylamino)quinolin-2-yl}-4-methoxy-benzamidein 10 mL of pyridine was stirred at room temperature with 0.2 mL of DBUand 150 mg of pyridinium-SO₃ komplex for 2 hours. The reaction mixturewas concentrated to dryness, the residue was chromatographed on silicagel using a mixture of ethyl acetate/methanol/25% ammonia =200/80/2 aseluent to give 10 mg of title compound as ammonium salt. LC-MS: MH⁺ 518;Ret. time: 6.22 minutes.

Example 13N-{4-(Benzylamino)-3-cyano-6-[formyl(methyl)amino]quinolin-2-yl}-4-methoxybenzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, R⁴ stands for Me, R⁵ stands for CHO and R⁶ for 4-methoxyphenyl,X for NH and the value of n is 1.

A suspension of 110 mg ofN-{4-(benzylamino)-3-cyano-6-(methylamino)quinolin-2-yl}-4-methoxybenzamidewas refluxed in 10 mL of ethyl formate for 24 hours. After cooling theprecipitated crystals were filtered off, washed with ethyl formate andethanol to give 30 mg of title compound, m.p.: 237-240° C. LC-MS: MH⁺465; Ret. time: 6.11 min.

Example 144-Methoxy-N-[6-(4-formylpiperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamide

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, group a.) for 4-formylpiperazino, R⁶ for 4-methoxyphenyl, X forNH and the value of n is 1.

A solution of 120 mg of4-methoxy-N-[6-(piperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamidein 5 mL of ethyl formate was refluxed for 2 hours. After cooling thesolvent was evaporated and the residue was recrystallysed from methanolto give 65 mg of the title compound. mp: 243° C.

¹H-NMR (DMSO-d₆) 10.6 ppm (s, 1H), 8.48 (m, 1H), 8.1 (s, 1H), 7.94 (m,2H), 7.6-7.1 (m, 8H), 7.05 (m, 2H), 5.1 (d, 2H), 3.82 (s, 3H), 3.7-3.2(m, 8H).

Example 154-[2-(Benzoylamino)-4-benzylamino)-3-cyanoquinolin-6-yl]piperazine-1-sulfonicacid

In the general formula (I) R¹ and R² stand for hydrogen atom, R³ forphenyl, group a.) for 4-sulfonylpiperazino, R⁶ for 4-methoxyphenyl, Xfor NH and the value of n is 1.

A solution of 50 mg of4-methoxy-N-[6-(piperazin-1-yl)-4-benzylamino-3-cyanoquinolin-2-yl]benzamidein 0.5 mL pyridine and pyridinium-SO₃ complex was refluxed for 2 hoursand working up according to a procedure published in the literature (G.F. Smith and D. A. Taylor, Tetrahedron, 29, 669, 1973).

LC-MS: MH⁺ 572; Ret. time: 5.9 min.

Structure and physical characteristics of further compounds of thegeneral formula (I), wherein R¹ and R² stand for hydrogen atom, X standsfor —NH— and n represents 1, are shown in Table I

TABLE I

LC-MS Mp MH⁺/Ret. Process of No R³ R⁴ R⁵ R⁶ (° C.) time(min) example 16

Me Me

211 4 17

Me Me

208 4 18

Me Me

198 4 19

Me Me

176 4 20

Me Me

152 4 21

Me Me

163 4 22

Me Me

172 4 23

Me Me

158 4 24

Me Me

170 4 25

Me Me

171 4 26

Me Me

165 4 27

Me Me

148 4 28

H H

235 424/5.65 7 29

H H

234 470/6.12 7 30

H H

222 444/5.54 7 31

H Me

160 444/5.74 9 32

H

217 517/5.86 11 33

H

257 509/5.70 11 34

H

208 523/5.97 11 35

H CHO

465/7.53 10 36

H CHO

452/5.69 10 37

H CHO

247 471/5.75 10 38

Me

523/6.14 12 39

Me CHO

248 471/5.86 13

Example 40

Tablets of the following composition are prepared by known methods:

Active ingredient: 25 mg Lactose 50 mg Avicel 21 mg Crospovidone  3 mgMagnesium stearate  1 mgPharmacological Testing

-   -   Biology    -   Methods    -   Human adenosine A₃ receptor binding

Preparing membrane suspension: ovarium cells of Chinese hamsterexpressing cloned human A₃ receptors (further: CHO-hA₃) areappropriately cultured and maintained. Achieving confluent cell layer,the medium is removed from the cells by washing them with 37° C. PBS,then the cell are suspended in ice cold PBS, centrifuged (1000×g 10 min)(Sigma 3K30) and homogenized using teflon homogenizer (B.Braun Potter S)at 1500/min rotation speed, for 15 sec. in the following buffer: 50 mMTris, 10 mM MgCl₂, 1 mM EDTA, pH 8.0. The suspension is centrifuged(43.000 g, 10 min). The pellet is suspended in the above buffer, proteinconcentration 0.1 mg/ml (Bradford method). Aliquots of the membranepreparation are stored at −80° C.

Binding protocol: incubate CHO-hA₃ membrane preparation (2 μg proteincontent) in incubation buffer (50 mM Tris, 10 mM MgCl₂, 1 mM EDTA, 3U/mL adenosine deaminase, pH 8.0), in the presence of 0.5 nM[¹²⁵I]AB-MECA (p-amino-3-iodo-benzyl-5′-N-methylcarboxamido-adenosine)(100.000 cpm) and 100 μM R-PIA (N⁶-[L-2-phenylisopropyl]adenosine) todefine non-specific binding of test compound in a total volume of 50 μLfor 1 hr at room temperature. Filter over Whatman GF/B glass fibrefilters (presoaked in 0.5% polyethylenimine for 3 hours), wash 4× with 1mL ice-cold 50 mM Tris, 10 mM MgCl₂, 1 mM EDTA (pH 8.0) on 96-wellBrandel Cell Harvester. Detection of activity: in gamma-counter (1470Wizard, Wallac). Inhibition [%]=100−((activity in the presence of testcompound−non-specific activity)/(total activity−non-specificactivity))*100

Human Adenosine A₁ Receptor Binding

Preparing membrane suspension: ovarium cells of Chinese hamsterexpressing cloned human A₁ receptors (further: CHO-hA₁) areappropriately cultured and maintained. Achieving confluent cell layer,the medium is removed from the cells by washing them with 37° C. PBS,then the cell are suspended in ice cold PBS, centrifuged (1000×g 10 min)(Sigma 3K30) and homogenized using teflon homogenizer (B.Braun Potter S)at 1500/min rotation speed, for 15 sec. in the following buffer: 50 mMTris, 10 mM HCl, pH 7.4. The suspension is centrifuged (43.000 g, 10min). The pellet is suspended in the above buffer, protein concentration5 mg/ml (Bradford method). Aliquots of the membrane preparation arestored at −80° C.

Binding protocol: incubate CHO-hA₁ membrane preparation (50 μg proteincontent) in incubation buffer (50 mM Tris, 3 U/mL adenosine deaminase,pH 7.4), 10 nM [³H]CCPA (2-chloro-N⁶-cyclopenthyl-adenosine) (80,000dpm) and 10 μM R-PIA (N⁶-[L-2-phenylisopropyl]adenosine) to define thenon-specific binding or test compound in a total volume of 100 μL for 3hr at room temperature. Filter over Whatman GF/B glass fibre filters(presoaked in 0.5% polyethylenimine for 3 hours), wash 4× with 1 mLice-cold 50 mM Tris (pH 7.4) on 96-well Brandel Cell Harvester.Detection of activity: in the presence of 200 μL of HiSafe-3 coctail inbeta-counter (1450 Microbeta, Wallac). Inhibition [%]=100−((activity inthe presence of test compound−non-specific activity)/(totalactivity−non-specific activity))*100

Human Adenosine A_(2a) Receptor Binding

Binding protocol: Incubate 7 μg of membranes (human A_(2a) adenosinereceptors transfected into HEK-293 cells, source: Receptor Biology,Inc.), buffer (50 mM Tris-HCl, 10 mM MgCl₂, 1 mM EDTA, 2 U/mL adenosinedeaminase, pH 7.4), 20 nM [³H]CGS-21680(2-[p-(2-carbonylethyl)-phenylethylamino]-5′-N-ethylcarboxamido-adenosine)(200.000 dpm) and 50 μM NECA (5′-N-ethylcarboxamido-adenosine) to definethe non-specific binding of test compound, in a total volume of 100 μlfor 90 min at room temperature. Filter in vacuum over Whatman GF/B glassfibre filters (presoaked for 3 hours in 0.5% polyethylenimine), wash 4×with 1 mL ice-cold 50 mM Tris, 10 mM MgCl₂, 1 mM EDTA, 0.9% NaCl, pH7.4) on 96-well Brandel Cell Harvester. Detection of activity: inbeta-counter (1450 Microbeta, Wallac) in the presence of 200 μL ofHiSafe-3 coctail. Inhibition [%]=100−((activity in the presence of testcompound−non-specific activity)/(total activity−non-specificactivity))*100.

Human adenosine A_(2b) Receptor Binding

Binding protocol: incubate 20.8 μg of membranes (human A_(2b) adenosinereceptors transfected into HEK-293 cells, source: Receptor Biology,Inc.), buffer (50 mM Tris-HCl, 10 mM MgCl₂, 1 mM EDTA, 0.1 mMbenzamidine, 2 U/mL adenosine deaminase, pH 6.5), 32.4 nM [³H]DPCPX(8-cyclopenthyl-1,3-dipropylxanthine) (800.000 dpm) and 100 μM NECA(5′-N-ethylcarboxamido-adenosine) to define non-specific binding or testcompound in a total volume of 100 μL for 30 min at room temperature.Filter under 25 mm Hg vacuum over Whatman GF/C glass fibre filters(presoaked in 0.5% polyethylenimine for 3 hours), wash 4× with 1 mLice-cold 50 mM Tris-HCl (pH 6.5) on 96-well Brandel Cell Harvester.Detection of activity: in the presence of 200 μL of HiSafe-3 coctail inbeta-counter (1450 Microbeta, Wallac). Inhibition [%]=100−((activity inthe presence of test compound−non-specific activity)/(totalactivity−non-specific activity))*100

Results

We consider the compounds as biologically active ones if they inhibitthe binding of the radioligand on human adenosine A₃ receptors with anactivity above 80% at 1 μM in our experimental conditions.

The dissociation constant (K_(d)) of [¹²⁵I]AB-MECA on CHO-hA₃ membranepreparation is determined by isotope saturation studies with the help ofScatchard analysis (G. Scatchard, Ann. N. Y. Acad. Sci. 51:660, 1949).The IC₅₀ is converted to an affinity constant (K_(i)) by application ofthe Cheng-Prusoff equation (Y. J. Cheng and W. H. Prusoff, Biochem.Pharmacol. 22:3099, 1973).

A number of the compounds of the general formula (I) displayedremarkable biological effects. The most active compounds of the generalformula (I) were those defined in claims 2-4. Especially advantageousare the compounds given in the Examples, their K_(i) values are in therange of 0.5 nM and 900 nM, preferably 0.5 nM and 700 nM. K_(i) valuesof the most advantageous compounds are in the range of 0.5 nM 18 nM,most preferably 0.5 and 15 nM.

The compounds possess good bioavailability and a selectivity of at least3 order of magnitude, in respect to the human adenosine A₁, A_(2a) andA_(2b) receptor subtypes.

Further, the duration of their action at intravenous and oraladministration is long, their ED₅₀ values are low, their toxicologicaland side-effect profiles are advantageous.

These above data are favourable for the therapeutic application of thecompound of the general formula (I).

1. A compound of formula (I),

wherein R¹ is hydrogen atom or a straight or branched C₁₋₄ alkyl; R² ishydrogen atom or a straight or branched C₁₋₄ alkyl; R³ is hydrogen atom,straight or branched C₁₋₄ alkyl, or C₃₋₆ cycloalkyl, phenyl, thienyl, orfuryl, each of which is optionally substituted with one or more straightor branched C₁₋₄ alkyl, straight or branched C₁₋₄ alkoxy, or halogenatom, or six- or five-membered heteroaromatic ring containing one, twoor three nitrogen atoms, or five-membered heteroaromatic ring containingone nitrogen atom and one oxygen atom, or one nitrogen atom and onesulphur atom, wherein the heteroaromatic ring is optionally substitutedwith one or more straight or branched C₁₋₄ alkyl, straight or branchedC₁₋₄ alkoxy, or halogen atom; R⁴ and R⁵ taken together with the nitrogenatom to which they are attached form a group of formula a.)

R⁶ is hydrogen atom, straight or branched C₁₋₄ alkyl, phenyl, benzyl,thienyl, or furyl, each of which is optionally substituted withmethylenedioxy, or one or more straight or branched C₁₋₄ alkyl, straightor branched C₁₋₄ alkoxy, hydroxy, trifluoromethyl, cyano or halogenatom, or six- or five-membered heteroaromatic ring containing one, twoor three nitrogen atoms, or one nitrogen atom and one oxygen atom, orone nitrogen atom and one sulphur atom, wherein the heteroaromatic ringis optionally substituted with one or more straight or branched C₁₋₄alkyl, straight or branched C₁₋₄ alkoxy or halogen atom; R⁷ and R⁸independently are hydrogen atom, straight or branched C₁₋₄ alkyl or C₃₋₆cycloalkyl; X is —CH₂—, —NH—, —NR⁹—, sulphur atom, oxygen atom, sulphoor sulphoxy; R⁹ is straight or branched C₁₋₄ alkyl or C₃₋₆ cycloalkyl; Zis—O—; n is zero, 1 or 2; m is 1, 2, or 3; o is 1, 2, or 3; p is zero or1; and r is zero or 1, or a salt, or tautomer, desmotrop or opticallyactive isomer thereof, or a salt of the tautomer, desmotrop or opticallyactive isomer.
 2. The compound according to claim 1, wherein R³ ishydrogen, straight or branched C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or phenyl,thienyl, or furyl, each of which is optionally substituted with one ormore straight or branched C₁₋₄ alkyl, straight or branched C₁₋₄ alkoxy,or halogen; a salt or tautomer, desmotrop or optically active isomerthereof, or a salt of the tautomer, desmotrop or optically activeisomer.
 3. The compound according to claim 1, wherein: R¹ is hydrogen ormethyl; R² is hydrogen or methyl; R³ is phenyl, thienyl, or furyl; R⁶stands for 4-methoxyphenyl, 3-methylphenyl, 3-methoxyphenyl,3,4-methylene-dioxypheny, 4-fluorophenyl, 2-thienyl or 2-furyl; R⁷ andR⁸ independently stand for hydrogen atom, straight or branched C₁₋₄alkyl or C₃₋₆ cycloalkyl; X is —NH—, or —O—; n is 1; m is 2; o is 2; pis 0; and r is 0, or a salt or tautomer, desmotrop or optically activeisomer thereof, or a salt of the tautomer, desmotrop or optically activeisomer.
 4. The compound according to claim 1, which is:4-Methoxy-N-[6-(morpholin-4-yl)-4-benzylamino-3-cyanoquinolin-2-yl]-benzamide,or a salt or tautomer, desmotrop or optically active isomer thereof, ora salt of the tautomer, desmotrop or optically active isomer.
 5. Aprocess for preparing the compound according to claim 1, comprisingselectively hydrolyzing a bis-carboxamide of formula (II″),

wherein R^(1″) is hydrogen atom or a straight or branched C₁₋₄ alkyl;R^(2″) is hydrogen atom or a straight or branched C₁₋₄ alkyl; R^(3″) ishydrogen atom, straight or branched C₁₋₄ alkyl, C₃₋₆ cycloalkyl, phenyl,thienyl, or furyl, each of which is optionally substituted by one ormore straight or branched C₁₋₄ alkyl, straight or branched C₁₄ alkoxy,or halogen atom, or six- or five-membered heteroaromatic ring containingone, two or three nitrogen atoms, or five-membered heteroaromatic ringcontaining one nitrogen atom and one oxygen atom, or one nitrogen atomand one sulphur atom, wherein the heteroaromatic ring is optionallysubstituted by one or more straight or branched C₁₋₄ alkyl, straight orbranched C₁₋₄ alkoxy, or halogen atom; R^(4″) and R^(5″) taken togetherwith the nitrogen atom to which they are attached form a nitro or agroup of formula a″.),

R^(6″) is hydrogen atom or straight or branched C₁₋₄ alkyl, phenyl,benzyl, thienyl, or furyl, each of which is optionally substituted withmethylenedioxy, or with one or more straight or branched C₁₋₄ alkyl,straight or branched C₁₋₄ alkoxy, hydroxy, trifluoromethyl, cyano orhalogen atom, or six- or five-membered heteroaromatic ring containingone, two or three nitrogen atoms, or one nitrogen atom and one oxygenatom, or one nitrogen atom and one sulphur atom, wherein theheteroaromatic ring is optionally substituted with one or more straightor branched C₁₋₄ alkyl, straight or branched C₁₋₄ alkoxy or halogenatom; R^(7″) and R^(8″) independently are hydrogen atom, straight orbranched C₁₋₄ alkyl or C₃₋₆ cycloalkyl; X″ is —CH₂—, —NH—, —NR^(9″),sulphur atom, oxygen atom, sulpho or sulphoxy; R^(9″) is straight orbranched C₁₋₄ alkyl or C₃₋₆ cycloalkyl; Z″ is —O—; n″ is zero, 1 or 2;m″ is 1, 2, or 3; o″ is 1, 2, or 3; p″ is zero or 1; and r″ is zero or1, and optionally removing a protective group.
 6. The process accordingto claim 5, wherein the selective hydrolysis is carried out using alkalihydroxide dissolved in alcoholic medium.
 7. A pharmaceutical compositioncomprising a pharmaceutically effective amount of one or more compoundsaccording to claim 1, or a salt or tautomer, desmotrop or opticallyactive isomer thereof, or a salt of the tautomer, desmotrop or opticallyactive isomer, and one or more auxiliary materials.
 8. A pharmaceuticalcomposition comprising a pharmaceutically effective amount of one ormore compounds according to claim 3, or a salt or tautomer, desmotrop oroptically active isomer thereof, or a salt of the tautomer, desmotrop oroptically active isomer, and one or more auxiliary materials.